Anti-allergen antibodies

ABSTRACT

The present invention generally relates to antibodies or binding fragments thereof capable of binding an allergen, in particular a food allergen as well as pharmaceutical compositions comprising such antibodies or binding fragments thereof for the treatment of allergy, in particular food allergy. In addition the invention relates to methods for evaluating the capacity of a candidate antibody or binding fragment thereof to inhibit allergen binding/and/or allergen- induced activity in a human and methods of detecting or quantifying whether an allergen is present in a sample.

FIELD OF THE INVENTION

The present invention generally relates to antibodies or bindingfragments thereof capable of binding an allergen, in particular a foodallergen as well as pharmaceutical compositions comprising suchantibodies or binding fragments thereof for the treatment of allergy, inparticular food allergy. In addition the invention relates to methodsfor evaluating the capacity of a candidate antibody or binding fragmentthereof to inhibit allergen binding/and/or allergen-induced activity ina human and methods of detecting or quantifying whether an allergen ispresent in a sample.

BACKGROUND OF THE INVENTION

Food allergies are reactions to ingested foods that may lead to clinicalmanifestations from skin, respiratory and gastrointestinal symptoms upto severe and life-threatening reactions, i.e. systemic anaphylaxis.Allergic reactions to many different types of foods have been described,but some of the most common are reactions to peanuts and shellfish. Suchimmediate hypersensitivity reactions are caused by immune responses toenvironmental antigens, also called allergens. Allergen encounterresults in the production of immunoglobulin E (IgE) antibodies byantibody-producing cells of the blood, the so-called B cells. IgEantibodies specific for the allergens bind to FcεRI receptors on mastcells and basophils, and upon critical food intake the allergens arebinding to pre-bound IgE on the surface of mast cells and DB:ADWbasophils. When these cell-associated IgE antibodies are cross-linked bythe allergen, mast cells and basophils are activated to rapidly releasea variety of mediators (e.g. histamine, leukotriene, lipid mediators),thereby triggering symptoms of allergy.

Systemic anaphylaxis, as manifested by urticaria, angioedema,bronchospasm, diarrhea, dysrhythmias, and cardiovascular collapse, isresponsible for a large number of emergency visits, hospital admissions,and deaths each year. Food allergy is responsible for 20-30% of theemergency visits for anaphylaxis and 100-200 deaths annually in theUnited States and 50-60% of these are caused by peanut allergy.

Indeed, allergy to peanuts is the most frequent cause of food-relateddeath. Compared to other food allergies, peanut allergy is less likelyto be outgrown. Individuals may be so sensitive to peanut allergens thatsevere systemic reactions can occur in response to minute contaminantsof the allergen introduced accidentally during food preparation.Accidental ingestions of peanuts may result in severe and fatalreactions and occur in up to 25-75% of patients over a 5-year period,causing significant anxiety in patients and in families of children withpeanut allergy.

Ara h 2 was described as the most important peanut allergen, as it wasidentified as a predictor of clinical reactivity to peanut.Polysensitization to Ara h 2 and Ara h 1 and/or Ara h 3 appeared to bepredictive of more severe reactions (Bublin, M. et al. Cross-reactivityof peanut allergens. Curr. Allergy Asthma Rep. 14, 426 (2014)). Inaddition, Ara h 6 has recently emerged as a major peanut allergen(Codreanu F, et al. A novel immunoassay using recombinant allergenssimplifies peanut allergy diagnosis. Int. Arch. Allergy Immunol. 154,216-226 (2011)). Ara h 1 (cupin) contributes to 12-16% of the totalprotein content of a peanut; Ara h 2 (2S albumin) to 5.9-9.3%. Together,Ara h 1, 2 and 3 account for 75% of total protein content. All knownpeanut allergen classes comprise 85% of the total peanut protein content(Bublin et al.). Mono-sensitization to a single peanut allergen is veryrare, but sensitization occurs with a high degree of heterogeneity to anumber of peanut allergens (Shreffler, W. G., et al. Microarrayimmunoassay: Association of clinical history, in vitro IgE function, andheterogeneity of allergenic peanut epitopes. J. Allergy Clin. Immunol.113, 776-782 (2004)).

Allergen immunotherapy, also known as desensitization, involves exposingthe patient to increasing amounts of allergen and is commonly used totreat patients allergic to many seasonal and perennial allergens. Incontrast, in peanut allergic patients allergen immunotherapy has notbeen established, primarily due to the safety risk of treating patientswith highly allergenic peanut proteins. In contrast to symptomaticmedication such as epinephrine, antihistamines, β-adrenergic agonist andcorticosteroid, effective allergen immunotherapy causes an increasedimmunological tolerance to the allergen or leads to a protectiveimmunity against allergens.

OBJECTIVES AND SUMMARY OF THE INVENTION

In order to meet the above needs, it is one objective of the inventionto provide antibodies or binding fragments thereof for the effectivetreatment of allergies, in particular food allergies, such as peanutallergy.

The antibodies or binding fragments thereof described herein may becapable of reducing, inhibiting or neutralizing allergen-mediatedbiological activity. In particular, the antibodies may be capable ofreducing or inhibiting the binding of an IgE antibody to the foodallergen. Thus, the antibodies or binding fragments thereof according tothe invention decrease or inhibit the activation of the mast cells orbasophils and therefore decrease or prevent the release of mediators(e.g. histamine, lipid mediators, leukotriene). Thereby, the antibodiesdescribed herein may inhibit allergy symptoms that would usually occurin the patient after contact with the allergen (e.g. contact with theeyes, nose or mouth or food uptake).

The inventors found that treatment of patients suffering from peanutallergy with human-derived antibodies could be highly effective. Sincehuman-derived antibodies underwent the natural “evolution” in the humanbody, these antibodies are highly effective for the treatment of apatient.

The food allergen may be a peanut allergen. In an specific embodimentthe peanut allergen is selected from the group consisting of Ara h 1,Ara h 2, Ara h 3, Ara h 4, Ara h 5, Ara h 6/7, Ara h 8, Ara h 9 and Arah 10/11. Preferably, the allergen is selected from the group consistingof Ara h 1, Ara h 2, Ara h 3 and Ara h 6, or a combination thereof.Typically, the peanut allergen is of peanut origin, recombinantlyexpressed or is a synthetic peanut peptide.

Typically, the antibody may be a monoclonal antibody. In someembodiments, the antibody may be a recombinant antibody.

The antibody may be an IgG or IgA antibody. IgG and IgA compete with IgEfor binding sites on the allergen and thereby prevent recognition ofallergens by IgE bound to Fcε receptors on the surface of mast cells andbasophils. This may include direct competition by binding to the sameepitope or competition through steric hindrance. Furthermore, IgGantibodies bound to the allergen can lead to cross-linking of Fcε andinhibitory FcγRIIB receptors, resulting in the decrease of effector cellactivity. Thereby the IgG and IgA antibodies or binding fragmentsthereof according to the invention can be used for the effectiveprevention or treatment of allergies.

In some embodiments, the variable regions, portions thereof or the CDRsare human-derived. For example, the variable regions, portions thereofor the CDRs are derived from an IgE antibody and grafted in a scaffoldof an IgG or IgA antibody. In one embodiment, the peptide sequence ofthe antibody or binding fragment thereof is identical or at least 60%identical to the sequence of the antibody extracted from the human. Thehuman from which the antibody is extracted may be a human suffering frompeanut allergy, a peanut-sensitized human without clinical relevantallergy, a human suffering from peanut allergy that underwentimmunotherapy, a human that has outgrown peanut allergy and a human ofunknown clinical history for peanut allergy.

In some embodiments, the antibody or binding fragment thereof has anEC50 of at most 270 ng/ml, preferably at most 70 ng/ml, at most 40ng/ml, at most 25 ng/ml, at most 15 ng/ml, at most 4.9 ng/ml, at most1.3 ng/ml for at least one of the peanuts allergens selected from thegroup consisting of Ara h 2, Ara h 1, Ara h 3 and Ara h 6. Preferably,the antibody has an EC50 of at most 10 ng/ml, preferably at most 7ng/ml, more preferably at most 4.8 ng/ml, most preferably at most 2.8ng/ml for peanut extract.

The antibodies described herein may be capable of reducing, inhibitingor neutralizing allergen-mediated biological activity. In particular,the antibodies may be capable of reducing or inhibiting the binding ofan IgE antibody to the food allergen.

In one embodiment, the antibody or binding fragment thereof comprises alight chain variable region and/or a heavy chain variable region,wherein the light chain variable region comprises a CDR1 set forth inSEQ ID No: 1 or sequences at least 65% identical thereto, a CDR2 setforth in SEQ ID No: 2 or sequences at least 65% identical thereto, aCDR3 set forth in SEQ ID No: 3 or sequences at least 65% identicalthereto; wherein the heavy chain variable region comprises a CDR1 setforth in SEQ ID No: 4 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 5 or sequences at least 65% identicalthereto, and a CDR3 set forth in SEQ ID No: 6 or sequences at least 65%identical thereto.

In another embodiment refers to an antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 15 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 16 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 17 or sequences at least 65%identical thereto; wherein the heavy chain variable region comprises aCDR1 set forth in SEQ ID No: 18 or sequences at least 65% identicalthereto, a CDR2 set forth in SEQ ID No: 19 or sequences at least 65%identical thereto, a CDR3 set forth in SEQ ID No: 20 or sequences atleast 65% identical thereto.

A further embodiment refers to an antibody or binding fragmentcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 29 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 30 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 31 or sequences at least 65%identical thereto; wherein the heavy chain variable region comprises aCDR1 set forth in SEQ ID No: 32 or sequences at least 65% identicalthereto, a CDR2 set forth in SEQ ID No: 33 or sequences at least 65%identical thereto, a CDR3 set forth in SEQ ID No: 34 or sequences atleast 65% identical thereto.

A further embodiment refers to an antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 43 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 44 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 45 or sequences at least 65%identical thereto; and/or wherein the heavy chain variable regioncomprises a CDR1 set forth in SEQ ID No: 46 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 47 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 48 orsequences at least 65% identical thereto.

A further embodiment refers to an antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 57 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 58 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 59 or sequences at least 65%identical thereto; and/or wherein the heavy chain variable regioncomprises a CDR1 set forth in SEQ ID No: 60 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 61 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 62 orsequences at least 65% identical thereto.

A further embodiment refers to an antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 71 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 72 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 72 or sequences at least 65%identical thereto; and/or wherein the heavy chain variable regioncomprises a CDR1 set forth in SEQ ID No: 74 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 75 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 76 orsequences at least 65% identical thereto.

The antibodies are described in more detail below.

Preferably, the antibody is selected from the group consisting of a fulllength antibody, multispecific antibody, a single chain Fv fragment(scFv), a F(ab′) fragment, a F(ab′)2 fragment, a F(ab)c fragment, asingle domain antibody fragment (sdAB) or a multispecific antibodyfragment.

Another aspect of the invention refers to an antibody or bindingfragment thereof which competes with an antibody as described herein forspecific binding to the allergen.

A further aspect refers to a polynucleotide encoding the antibody orbinding fragment thereof as described herein. Another embodiment refersto a vector comprising a polynucleotide encoding the antibody or bindingfragment thereof as described herein. Another embodiment refers to acell comprising a polynucleotide encoding the antibody or bindingfragment thereof as described herein or a vector comprising apolynucleotide encoding the antibody or binding fragment thereof asdescribed.

A further aspect refers to a method for preparing an anti-allergenantibody or allergen-binding fragment thereof, consisting of culturingthe cell comprising a polynucleotide encoding the antibody or bindingfragment thereof as described herein and isolating the antibody orallergen binding fragment thereof from the cell or culture medium of thecell.

Moreover, the invention relates to an antibody composition comprising atleast two antibodies, wherein at least one of the antibodies is selectedfrom the antibodies as defined herein. In one embodiment the antibodycomposition comprises at least two of the antibodies selected from theantibodies as defined herein. In one embodiment the antibody compositioncomprises at least three of the antibodies selected from the antibodiesas defined herein.

Another aspect of the invention refers to a pharmaceutical compositioncomprising at least one of the compounds selected from the groupconsisting of antibody or binding fragment as described herein, theantibody composition as described herein, the polynucleotide asdescribed herein, the vector as described herein and the cell asdescribed herein.

In one embodiment, the pharmaceutical comprises the antibody or bindingfragment of any as described herein or the antibody composition asdescribed herein.

Another embodiment the pharmaceutical composition further comprises anadditional agent useful for treating peanut allergy. The additionalagent useful for treating peanut allergy may be a β-adrenergic agonist(e.g. epinephrine), antihistamine, corticosteroid, anti-IgE antibody,anti-IgE antibody binding fragment, peptide vaccine and furtherantibodies capable of binding to a peanut allergen. Preferably, thepharmaceutical composition comprises epinephrine.

Typically, the pharmaceutical composition may comprise a pharmaceuticalacceptable carrier.

Preferably, the pharmaceutical composition is intended for subcutaneous,intravenous, intramuscular, intraperitoneal, intranasal and/orinhalative administration. The antibody or binding fragment thereof, inparticular the pharmaceutical composition comprising the antibody orbinding fragment, may be administered once or more times.

Other aspects of the invention refer to the antibodies or bindingfragments, antibody composition, polynucleotide, vector, cell orpharmaceutical composition as described herein for use in the treatmentof allergy. The allergy may be a food allergy, such as peanut allergy.In particular, the invention refers to the antibodies or bindingfragments as described herein, the antibody composition as describedherein and the pharmaceutical composition as described herein for use inthe treatment of food allergy, such as peanut allergy. The treatment maybe prophylactic or therapeutic.

Another aspect refers to a method of evaluating the capacity of acandidate antibody or binding fragment thereof to inhibit allergenbinding/and/or allergen-induced activity in a human,

wherein the method comprises

(i) incubating the candidate antibody or binding fragment thereof with acomposition comprising IgEs derived from the human and a food allergen;

(ii) evaluating whether the candidate antibody inhibits allergenbinding/and/or allergen-induced activity in the composition comprisingIgEs derived from the human.

Thus, the method of the invention allows to tailor the therapy to thepatient based on the predicted response to the antibodies or fragmentsthereof according to the invention. This approach is also referred as“personalized medicine” approach thereby allows to identify the mosteffective antibody or antibody combination.

In one embodiment the method further comprises the step of

(iii) determining whether the administration of the candidate antibodyis a suitable treatment for a patient suffering from food allergy basedon the result of step (ii).

In other words, the present application refers to methods fordetermining whether a patient suffering from food allergy is responsiveto a candidate antibody, comprising the steps of

(i) incubating the candidate antibody or binding fragment thereof with acomposition comprising IgEs derived from the human and a food allergen;

(ii) evaluating whether the candidate antibody inhibits allergen bindingand/or allergen-induced activity in the composition comprising IgEsderived from the human;

(iii) determining whether the administration of the candidate antibodyis a suitable treatment for the patient suffering from food allergybased on the result of step (ii).

Preferably the candidate antibody is an antibody of the invention asdescribed herein.

Preferably, in step (i) the composition comprising IgEs from theallergic patient comprises basophils. Typically, the basophils arederived from the patient. Alternatively, the basophils are donor-derivedIgE-stripped basophils.

In one embodiment, basophils are identified based on the lowside-scattered light (SSC) and the expression of CCR3. In addition,activated basophils are identified by measuring surface expression ofCD63 and/or surface expression of CD203c. Typically, in activatedbasophils the surface expression of CD63 and/or CD203c is high.

In another embodiment, the secretion of a mediator from leukocytes, inparticular basophils, is measured. In a preferred embodiment themediator is leukotriene, such as sulfidoleukotriene.

The measurement of secreted mediators, in particular leukotriene isadvantageous, since the measurement of the mediators, preferablyleukotriene, released from the leukocytes, in particular basophils, canbe detected by ELISA and therefore can be easily carried out inhigh-throughput format. Leukotriene is released from the basophils in adose dependent manner. Thus, it is particularly suitable for highlysensitive assays.

The composition comprising IgEs derived from the human may be plasma,sera, blood, saliva, peripheral blood mononuclear cell (PBMC),leukocytes, basophils or IgEs stripped from basophils.

When measuring secreted mediators, preferably leukotriene, leukocytesmay be used as composition comprising IgEs. This makes the assaystraight forward, since it is not necessary to distinguish or isolatethe basophils from the other cell fractions (such as neutrophils,eosinophils, lymphocytes and monocytes) of the leukocyte pool formeasuring secreted mediators.

Another aspect of the invention refers to a method of detecting orquantifying whether an allergen is present in a sample comprising thefollowing steps:

i) incubation of the sample with an antibody of the invention asdescribed herein or with an antibody composition of the invention asdescribed herein,

ii) detecting the antibody which is bound to allergen in the sample.

The antibody may be detectably labeled. Alternatively, the antibody isused with a second antibody that is detectably labeled. Preferably, theantibody is unlabeled and used in combination with a second antibodythat is detectably labeled. The detectable label may be selected fromthe group consisting of an enzyme, a radioisotope, a fluorophore, apeptide and a heavy metal.

FIGURE LEGENDS

FIG. 1. histogram distribution of the 84 allergic patients enrolled inthe clinical trial described in example 1, total IgG, IgG4, IgE antibodyreactivities against major peanut allergens (Ara h 1, 2 and 3).Seroreactivities are expressed as titer, which is defined as OD450 valueover background.

FIG. 2. histogram distribution of the 84 allergic patients enrolled inthe clinical trial described in example 1, total IgG, IgG4, IgE antibodyreactivities against major peanut allergen Ara h 6 and peanut extract.Seroreactivities are expressed as titer, which is defined as OD450 valueover background.

FIG. 3. EC50 ELISA determination of binding of exemplary anti-Ara h 2antibody 32B10 to naturally extracted peanut allergens (nAra h 1, nAra h2, nAra h 3, nAra h 6, Indoor Biotechnologies, Cardiff, UK),recombinantly expressed Ara h 2 (rAra h 2, Indoor Biotechnologies,Cardiff, UK) and natural peanut extract. Bovine serum albumin (BSA)serves as control. Similar binding was detected for the antibodies 37D5,12G10, 4B2, 2F8 and 7G6.

FIG. 4. Human-derived anti-peanut allergen monoclonal antibodies inhibitallergen-mediated activation of basophils derived from allergicpatients. Basophils were either left untreated or stimulated with peanutextract in the presence of human-derived antibodies or an isotypecontrol. A. Bars indicate activation of basophils expressed as % CD63+cells (mean±s.d, n=3). Pre-incubation of peanut extract with exemplaryantibodies (32B10, 37D5, 12G10, 4B2) or a combination of them (MY006cocktail) decreased basophil degranulation, whereas isotype control didnot have any effect. White bars: no addition of antibody.

FIG. 5. Human-derived anti-peanut allergen monoclonal antibodies inhibitallergen-mediated release of leukotriene (sLT) from leukocytes derivedfrom allergic patients. Leukocytes were stimulated with increasingconcentrations of peanut extract (PE; 0-30 ng/ml) in the presence of acocktail of human-derived antibodies (MY006) or an isotype control.Pre-incubation of peanut extract with MY006 cocktail inhibitedleukotriene release (curve shift to the right), whereas isotype controldid not have any effect.

FIG. 6. Effect of different anti-peanut allergen antibody combinationscan be evaluated by leukotriene (sLT) release assay. Leukocytes fromhealthy blood donors were isolated and surface IgE was removed byincubation with lactic acid. Leukocytes were re-sensitized with IgE byincubation with plasma from allergic patients. Re-sensitized leukocyteswere stimulated with peanut extract (0-30 ng/ml) in the presence ofdifferent human-derived monoclonal anti-peanut allergen antibodies or anisotype control. Pre-incubation of peanut extract (PE) withhuman-derived anti-peanut allergen antibodies inhibited leukotrienerelease to different extents, whereas isotype control did not have anyeffect.

DETAILED DESCRIPTION OF THE INVENTION

Before the invention is described in detail with respect to some of itspreferred embodiments, the following general definitions are provided.

The present invention as illustratively described in the following maysuitably be practiced in the absence of any element or elements,limitation or limitations, not specifically disclosed herein.

The present invention will be described with respect to particularembodiments and with reference to certain figures but the invention isnot limited thereto but only by the claims.

Where the term “comprising” is used in the present description andclaims, it does not exclude other elements. For the purposes of thepresent invention, the term “consisting of” is considered to be apreferred embodiment of the term “comprising of”. If hereinafter a groupis defined to comprise at least a certain number of embodiments, this isalso to be understood to disclose a group which preferably consists onlyof these embodiments.

For the purposes of the present invention, the term “obtained” isconsidered to be a preferred embodiment of the term “obtainable”. Ifhereinafter e.g. an antibody is defined to be obtainable from a specificsource, this is also to be understood to disclose an antibody which isobtained from this source.

Where an indefinite or definite article is used when referring to asingular noun, e.g. “a”, “an” or “the”, this includes a plural of thatnoun unless something else is specifically stated. The terms “about” or“approximately” in the context of the present invention denote aninterval of accuracy that the person skilled in the art will understandto still ensure the technical effect of the feature in question. Theterm typically indicates deviation from the indicated numerical value of±10%, and preferably of ±5%.

Technical terms are used by their common sense. If a specific meaning isconveyed to certain terms, definitions of terms will be given in thefollowing in the context of which the terms are used.

A first aspect relates to an antibody or binding fragment thereofcapable of binding to a food allergen.

The term “capable of binding to a food allergen” as used herein meansthat the antibody is specific for the allergen, i.e. it recognizes theallergen. Preferably, the antibody has a higher affinity for thespecific allergen, i.e. the specific food allergen, such as the peanutallergen Ara h 1, Ara h 2, Ara h 3, Ara h 6 than for other allergens.

The affinity or avidity of an antibody for an antigen can be determinedexperimentally using any suitable method; see, for example, Berzofsky etal., “Antibody-Antigen Interactions” In Fundamental Immunology, Paul, W.E., Ed., Raven Press New York, N Y (1984), Kuby, Janis Immunology, W. H.Freeman and Company New York, N Y (1992), and methods described herein.The measured affinity of a particular antibody-antigen interaction canvary if measured under different conditions, e.g., salt concentration,pH. Thus, measurements of affinity and other antigen-binding parameters,e.g., KD, IC50, EC50 are preferably made with standardized solutions ofantibody and antigen, and a standardized buffer.

Preferably the antibody or binding fragment thereof is human-derived.

The CDRs, the variable and/or the constant regions may stem from thesame human-derived antibody. Alternatively, the variable regions,portions thereof or the CDRs are human-derived and are grafted in anantibody framework. This antibody framework may be of human origin.Typically, the human-derived portions of the variable regions that aregrafted into the antibody framework comprise the CDRs.

The antibody or binding fragment thereof may comprise a CL and/or CHconstant region comprising an amino acid sequence selected from the CLamino acid sequences SEQ ID NOS: 9, 23, 37, 51 and 65 or an amino acidsequence with at least 60% identity and an amino acid sequence selectedfrom the CH amino acid sequences SEQ ID NOS: 10, 24, 38, 52 and 66 or anamino acid sequence with at least 60% identity.

The term “allergen” in the context of the invention is a compound thatis recognized by the immune system as foreign so that an immunoreactionto the allergen is evoked. In other words it refers to molecules thatare not present in the human body and may produce an abnormal immuneresponse by the activation of mast cells and basophils triggeringsymptoms of allergy. The skilled person understands that in the contextof the invention “autoantigens”, i.e. molecules that are produced by thehuman body are not considered as allergen.

The antibody may be capable of binding a food allergen. Food allergenmay be any food that that causes allergy or ingredient thereof, inparticular protein. The food allergen may be for example milk, eggs,crustacean, shellfish, tree nuts, peanuts, wheat or soybean oringredients thereof. In one embodiment the food allergen is peanut ortree nut. In a specific embodiment the food allergen is a legume, suchas soybean or peanut. Patients with peanut allergy may have a greaterchance of being allergic to other legumes such as soy.

In one embodiment, the allergen is a peanut allergen. The peanutallergen may be untreated or treated peanut, such as roasted peanut. Thepeanut allergen may be a peanut protein, such as Ara h 1, Ara h 2, Ara h3, Ara h 4, Ara h 5, Ara h 6/7, Ara h 8, Ara h 9 and Ara h 10/11. In aspecific embodiment the peanut protein may be in particular selectedfrom Ara h 1, Ara h 2, Ara h 3 and Ara h 6, or a combination thereof.The peanut allergen may be of peanut origin, recombinantly expressed oris a synthetic peanut peptide.

Typically, the antibody is a monoclonal antibody. In some embodiments,the antibody is a recombinant antibody.

Antibody or binding fragment thereof according to any one of thepreceding claims, wherein the antibody is an IgG, such as an IgG1, IgG2,IgG3 or IgG4 antibody, or IgA antibody.

In one embodiment the IgG may be an IgG1, IgG2 or IgG4 antibody.

The term “human-derived” in the context of the present invention meansthat at least the CDRs are derived from a human antibody. The“human-derived” antibody may contain further elements that are derivedfrom the human antibody, such as parts or the complete framework of theheavy and/or light chain variable regions and/or the parts or thecomplete of the heavy and/or light chain constant region. For example,the variable regions, portions thereof or the CDRs may be derived froman IgE antibody and grafted in a scaffold of an IgG or IgA antibody. Thepeptide sequence of the human-derived antibody may be at least 60%, atleast 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95% or at least 97% identical to the sequence of theantibody extracted from the human. Preferably, the peptide sequence ofthe human-derived antibody may be at least 90%, at least 95% , at least97%, at least 98% or at least 99% identical to the sequence of theantibody extracted from the human.

The human from which the antibody is extracted may be selected from thegroup of a human suffering from peanut allergy, a peanut-sensitizedhuman without clinical relevant allergy, a human suffering from peanutallergy that underwent immunotherapy, a human that has outgrown peanutallergy and a human of unknown clinical history for peanut allergy.

The antibodies of the invention may be used for the prevention ortreatment of allergy.

Allergy symptoms may be skin rash, itching skin, itching or tinglingsensation in or around the mouth or throat, headache, sneezing,swelling, nausea, diarrhea or anaphylaxis.

In some embodiments, the antibody or binding fragment thereof has anEC50 of at most 270 ng/ml, preferably at most 70 ng/ml, at most 40ng/ml, at most 25 ng/ml, at most 15 ng/ml, at most 4.9 ng/ml, at most1.3 ng/ml for at least one of the peanut allergens selected from thegroup consisting of Ara h 2, Ara h 1, Ara h 3 and Ara h 6. Preferably,the antibody has an EC50 of at most 10 ng/ml, preferably at most 7ng/ml, more preferably at most 4.8 ng/ml, most preferably 2.8 ng/ml forpeanut extract. The EC50 may be measured by an ELISA assay as describedherein.

In one embodiment, the antibody or binding fragment thereof comprises alight chain variable region and/or a heavy chain variable region,wherein the light chain variable region comprises a CDR1 set forth inSEQ ID No: 1 or sequences at least 65% identical thereto, a CDR2 setforth in SEQ ID No: 2 or sequences at least 65% identical thereto, aCDR3 set forth in SEQ ID No: 3 or sequences at least 65% identicalthereto; wherein the heavy chain variable region comprises a CDR1 setforth in SEQ ID No: 4 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 5 or sequences at least 65% identicalthereto, and a CDR3 set forth in SEQ ID No: 6 or sequences at least 65%identical thereto.

Another embodiment refers to an antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 15 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 16 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 17 or sequences at least 65%identical thereto; wherein the heavy chain variable region comprises aCDR1 set forth in SEQ ID No: 18 or sequences at least 65% identicalthereto, a CDR2 set forth in SEQ ID No: 19 or sequences at least 65%identical thereto, a CDR3 set forth in SEQ ID No: 20 or sequences atleast 65% identical thereto.

A further embodiment refers to an antibody or binding fragmentcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 29 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 30 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 31 or sequences at least 65%identical thereto; wherein the heavy chain variable region comprises aCDR1 set forth in SEQ ID No: 32 or sequences at least 65% identicalthereto, a CDR2 set forth in SEQ ID No: 33 or sequences at least 65%identical thereto, a CDR3 set forth in SEQ ID No: 34 or sequences atleast 65% identical thereto.

A further embodiment refers to an antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 43 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 44 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 45 or sequences at least 65%identical thereto; and/or wherein the heavy chain variable regioncomprises a CDR1 set forth in SEQ ID No: 46 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 47 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 48 orsequences at least 65% identical thereto.

A further embodiment refers to an antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 43 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 44 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 45 or sequences at least 65%identical thereto; and/or wherein the heavy chain variable regioncomprises a CDR1 set forth in SEQ ID No: 46 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 47 or sequences atleast 75% identical thereto, a CDR3 set forth in SEQ ID No: 48 orsequences at least 65% identical thereto.

A further embodiment refers to an antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 57 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 58 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 59 or sequences at least 65%identical thereto; and/or wherein the heavy chain variable regioncomprises a CDR1 set forth in SEQ ID No: 60 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 61 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 62 orsequences at least 65% identical thereto.

A further embodiment refers to an antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 71 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 72 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 73 or sequences at least 65%identical thereto; and/or wherein the heavy chain variable regioncomprises a CDR1 set forth in SEQ ID No: 74 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 75 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 76 orsequences at least 65% identical thereto.

One embodiment refers to an antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 1 or sequences at least 91% identical thereto, aCDR2 set forth in SEQ ID No: 2 or sequences at least 71% identicalthereto, a CDR3 set forth in SEQ ID No: 3; wherein the heavy chainvariable region comprises a CDR1 set forth in SEQ ID No: 4 or sequencesat least 80% identical thereto, a CDR2 set forth in SEQ ID No: 5 orsequences at least 82% identical thereto, a CDR3 set forth in SEQ ID No:6 or a sequence at least 94% identical thereto.

A further embodiment refers to an antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 71 or sequences at least 91% identical thereto, aCDR2 set forth in SEQ ID No: 72 or sequences at least 71% identicalthereto, a CDR3 set forth in SEQ ID No: 73; wherein the heavy chainvariable region comprises a CDR1 set forth in SEQ ID No: 74 or sequencesat least 80% identical thereto, a CDR2 set forth in SEQ ID No: 75 orsequences at least 82% identical thereto, a CDR3 set forth in SEQ ID No:76 or sequences at least 94% identical thereto.

A further embodiment refers to an antibody or binding fragment thereof,comprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a sequence setforth in SEQ ID No: 7 or sequences at least 70% identical thereto,and/or wherein the heavy chain variable region comprises a sequence setforth in SEQ ID No: 8 or sequences at least 70% identical thereto.

A further embodiment refers to an antibody or binding fragment thereof,comprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a sequence setforth in SEQ ID No: 21 or sequences at least 70% identical thereto,and/or wherein the heavy chain variable region comprises a sequence setforth in SEQ ID No: 22 or sequences at least 70% identical thereto.

A further embodiment refers to an antibody or binding fragment thereof,comprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a sequence setforth in SEQ ID No: 35 or sequences at least 70% identical thereto,and/or wherein the heavy chain variable region comprises a sequence setforth in SEQ ID No: 36 or sequences at least 70% identical thereto.

A further embodiment refers to an antibody or binding fragment thereof,comprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a sequence setforth in SEQ ID No: 49 or sequences at least 70% identical thereto,and/or wherein the heavy chain variable region comprises a sequence setforth in SEQ ID No: 50 or sequences at least 70% identical thereto.

A further embodiment refers to an antibody or binding fragment thereof,comprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a sequence setforth in SEQ ID No: 63 or sequences at least 70% identical thereto,and/or wherein the heavy chain variable region comprises a sequence setforth in SEQ ID No: 64 or sequences at least 70% identical thereto.

A further embodiment refers to an antibody or binding fragment thereof,comprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a sequence setforth in SEQ ID No: 77 or sequences at least 70% identical thereto,and/or wherein the heavy chain variable region comprises a sequence setforth in SEQ ID No: 78 or sequences at least 70% identical thereto.

A further embodiment refers to an antibody or binding fragment thereof,comprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a sequence setforth in SEQ ID No: 7 or sequences at least 91% identical thereto,and/or wherein the heavy chain variable region comprises a sequence setforth in SEQ ID No: 8 or sequences at least 88% identical thereto.

A further embodiment refers to an antibody or binding fragment thereof,comprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a sequence setforth in SEQ ID No: 77 or sequences at least 91% identical thereto,and/or wherein the heavy chain variable region comprises a sequence setforth in SEQ ID No: 78 or sequences at least 88% identical thereto.

A further embodiment refers to an antibody or binding fragment thereof,comprising in its variable region at least one CDR of the light chainvariable region comprising the amino acid sequence SEQ ID NO: 7 and/orthe heavy chain variable region comprising the amino acid sequence SEQID NO: 8.

A further embodiment refers to an antibody or binding fragment thereof,comprising in its variable region the CDR1, CDR2 and CDR3 of the lightchain variable region comprising the amino acid sequence SEQ ID NO: 7and the heavy chain variable region comprising the amino acid sequenceSEQ ID NO: 8.

A further embodiment refers to an antibody or binding fragment thereof,comprising in its variable region at least one CDR of the light chainvariable region comprising the amino acid sequence SEQ ID NO: 21 and/orthe heavy chain variable region comprising the amino acid sequence SEQID NO: 22.

A further embodiment refers to an antibody or binding fragment thereof,comprising in its variable region the CDR1, CDR2 and CDR3 of the lightchain variable region comprising the amino acid sequence SEQ ID NO: 21and the heavy chain variable region comprising the amino acid sequenceSEQ ID NO: 22.

A further embodiment refers to an antibody or binding fragment thereof,comprising in its variable region at least one CDR of the light chainvariable region comprising the amino acid sequence SEQ ID NO: 35 and/orthe heavy chain variable region comprising the amino acid sequence SEQID NO: 36.

A further embodiment refers to an antibody or binding fragment thereof,comprising in its variable region the CDR1, CDR2 and CDR3 of the lightchain variable region comprising the amino acid sequence SEQ ID NO: 35and the heavy chain variable region comprising the amino acid sequenceSEQ ID NO: 36.

A further embodiment refers to an antibody or binding fragment thereof,comprising in its variable region at least one CDR of the light chainvariable region comprising the amino acid sequence SEQ ID NO: 49 and/orthe heavy chain variable region comprising the amino acid sequence SEQID NO: 50.

A further embodiment refers to an antibody or binding fragment thereof,comprising in its variable region the CDR1, CDR2 and CDR3 of the lightchain variable region comprising the amino acid sequence SEQ ID NO: 49and the heavy chain variable region comprising the amino acid sequenceSEQ ID NO: 50.

A further embodiment refers to an antibody or binding fragment thereof,comprising in its variable region at least one CDR of the light chainvariable region comprising the amino acid sequence SEQ ID NO: 63 and/orthe heavy chain variable region comprising the amino acid sequence SEQID NO: 64.

A further embodiment refers to an antibody or binding fragment thereof,comprising in its variable region the CDR1, CDR2 and CDR3 of the lightchain variable region comprising the amino acid sequence SEQ ID NO: 63and the heavy chain variable region comprising the amino acid sequenceSEQ ID NO: 64.

A further embodiment refers to an antibody or binding fragment thereof,comprising in its variable region at least one CDR of the light chainvariable region comprising the amino acid sequence SEQ ID NO: 77 and/orthe heavy chain variable region comprising the amino acid sequence SEQID NO: 78.

A further embodiment refers to an antibody or binding fragment thereof,comprising in its variable region the CDR1, CDR2 and CDR3 of the lightchain variable region comprising the amino acid sequence SEQ ID NO: 77and the heavy chain variable region comprising the amino acid sequenceSEQ ID NO: 78.

The person skilled in the art knows that each variable domain (the heavychain VH and light chain VL) of an antibody comprises threehypervariable regions, sometimes called complementarity determiningregions or “CDRs” flanked by four relatively conserved framework regionsor “FRs” and refer to the amino acid residues of an antibody which areresponsible for antigen-binding. Exemplary conventions that can be usedto identify the boundaries of CDRs include, e.g., the Kabat definition,the Chothia definition, and the AbM definition. In general terms, theKabat definition is based on sequence variability, the Chothiadefinition is based on the location of the structural loop regions, andthe AbM definition is a compromise between the Kabat and Chothiaapproaches. See, e.g., Kabat, “Sequences of Proteins of ImmunologicalInterest,” National Institutes of Health, Bethesda, Md. (1991);Al-Lazikani et al., (1997), J. Mol. Biol. 273:927-948; and Martin etal., (1989), Proc. Natl. Acad. Sci. USA 86:9268-9272. For example, thehypervariable regions or CDRs of the human IgG subclass of antibodycomprise amino acid residues from residues 24-34 (L1), 50-56 (L2) and89-97 (L3) in the light chain variable domain and 31-35 (H1), 50-65 (H2)and 95-102 (H3) in the heavy chain variable domain as described by Kabatet al., and/or those residues from a hypervariable loop, i.e. residues26-32 (L1), 50-52 (L2) and 91-96 (L3) in the light chain variable domainand 26-32 (H1), 53-55 (H2) and 96-101 (H3) in the heavy chain variabledomain as described by Chothia et al., J. Mol. Biol. 196 (1987),901-917. Framework or FR residues are those variable domain residuesother than and bracketing the hypervariable regions. Accordingly, theterm “CDR” refers to the complementarity determining region orhypervariable region amino acid residues of an antibody that participatein or are responsible for antigen-binding. The CDRs as described hereinare defined according to Kabat et al. (1991) as described in Sequencesof Proteins of Immunological Interest.

Binding fragments may thus include portions of an intact full lengthantibody, such as an antigen binding or variable region of the completeantibody. Examples of antibody fragments include F(ab′), F(ab′)2,F(ab)c, and Fv fragments; diabodies; linear antibodies; single-Fvfragments (e.g., scFv); multispecific antibody fragments such asbispecific, trispecific, and multispecific antibodies (e.g., diabodies,triabodies, tetrabodies); minibodies; chelating recombinant antibodies;tribodies or bibodies; intrabodies; nanobodies; small modularimmunopharmaceuticals (SMIP), binding-domain immunoglobulin fusionproteins; camelized antibodies; VHH containing antibodies; chimericantigen receptor (CAR); and any other polypeptides formed from antibodyfragments. The skilled person is aware that the antigen-binding functionof an antibody can be performed by fragments of a full-length antibody.The binding fragment may have a length of at least 5, at least 8, atleast 10, at least 11, at least 12, at least 13, at least 14, at least15 or more amino acids. The fragment may have the antigen-bindingfunction of the antibody.

The skilled person is aware that also binding molecules capable ofbinding to a food allergen, in particular a peanut allergen and epitopesthereof, as defined herein, are encompassed by the invention. Inparticular encompassed are binding molecules capable of binding to afood allergen as defined herein and capable of preventing recognition offood allergens by IgE bound to FCE receptors. Hence the invention alsoincludes all aspects relating to these binding molecules, such aspharmaceutical compositions, kits, therapeutic and diagnostic uses aswell as methods relating to these binding molecules. Thus, the term“binding molecule” includes not only antibodies and binding fragmentsthereof but also includes non-antibody protein scaffold drugs, such asDARPins (for example reviewed in “Challenges and opportunities fornon-antibody scaffold drugs”, Vazquez-Lombardi R. et al Drug DiscoveryToday 20(10): 1271-1283) and other molecules that are not related topeptide structures.

The term “antibody” and “immunoglobulin” are used interchangeablyherein. Preferably, the antibody is a full length antibody ,multispecific antibody, a single chain Fv fragment (scFv), a F(ab′)fragment, a F(ab′)2 fragment, a F(ab)c fragment, a single domainantibody fragment (sdAB) or a multispecific antibody fragment.

A Fab fragment consists of the V_(L), V_(H), C_(L) and C_(H)1 domains.An F(ab′)2 fragment comprises two Fab fragments linked by a disulfidebridge at the hinge region. An Fd is the V_(H) and C_(H)1 domains of asingle arm of an antibody. An Fv fragment is the V_(L) and V_(H) domainsof a single arm of an antibody. An F(ab′)c fragment comprises two F(ab′)fragments plus part of the Fc domain. It is generated e.g. by plasmindigestion.

Binding fragments also encompass monovalent or multivalent, or monomericor multimeric (e.g. tetrameric), CDR-derived binding domains.

A bispecific antibody comprises two different binding specificities andthus binds to two different antigens or two different epitopes of thesame antigen. In one embodiment, the bispecific antibody comprises afirst antigen recognition domain that binds to a first antigen and asecond antigen recognition domain that binds to a second antigen. Inanother embodiment the bispecific antibody comprises a first antigenrecognition domain that binds to a first epitope of the antigen and asecond antigen recognition domain that binds to a second epitope of theantigen.

The determination of percent identity between multiple sequences ispreferably accomplished using the AlignX application of the Vector NTIAdvance™ 10 program (Invitrogen Corporation, Carlsbad Calif., USA). Thisprogram uses a modified Clustal W algorithm (Thompson et al., 1994. NuclAcids Res. 22:pp. 4673-4680; Invitrogen Corporation; Vector NTI Advance™10 DNA and protein sequence analysis software. User's Manual, 2004,pp.389-662). The determination of percent identity is performed with thestandard parameters of the AlignX application.

In these embodiments the sequence identity may be at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95% or at least 97%. Preferably, in all these embodiments the sequenceidentity is at least about 85%, more preferably at least about 90%, evenmore preferably at least about 95% and most preferably at least about98% or about 99%. Sequence identity may be determined over the wholelength of the respective sequences. Another aspect of the inventionrefers to an antibody or binding fragment thereof which competes with anantibody as described herein for specific binding to the allergen. Theskilled person is aware of methods for testing the capability of theantibody or binding fragment thereof to compete for specific binding tothe allergen, such as the competition ELISA assay as described elsewhereherein.

Some embodiments refer to an antibody which is capable of binding to atleast one of the Ara h 2 epitopes selected from the group consisting ofthe amino acid sequences set forth in SEQ ID NOs: 84, 88, 89, 90 and 91or fragments thereof.

A particular embodiment refers to an antibody which is capable ofbinding to the Ara h 2 epitope defined by the amino acid sequence setforth in SEQ ID NO: 84 or fragments thereof.

Another embodiment refers to an antibody which is capable of binding toat least one of the Ara h 2 epitopes selected from the group consistingof the amino acid sequences set forth in SEQ ID NOs: 88, 89, 90 and 91or fragments thereof.

The fragment of the epitope may comprise at least 5, at least 8, atleast 10, at least 11, at least 12, at least 13, at least 14, at least15, at least 17 or more amino acids.

Another embodiment refers to an antibody which is capable of binding allof the Ara h 2 epitopes selected from the group consisting of the aminoacid sequences set forth in SEQ ID NOs: 88, 89, 90 and 91.

Some embodiments refer to an antibody which is capable of binding to atleast one of the Ara h 2 epitopes selected from the group consisting ofthe amino acid sequences set forth in SEQ ID NOs: 84, 88, 89, 90 and 91.

A particular embodiment refers to an antibody which is capable ofbinding to the Ara h 2 epitope defined by the amino acid sequence setforth in SEQ ID NO: 84.

Another embodiment refers to an antibody which is capable of binding toat least one of the Ara h 2 epitopes selected from the group consistingof the amino acid sequences set forth in SEQ ID NOs: 88, 89, 90 and 91.

Another embodiment refers to an antibody which is capable of binding toall of the Ara h 2 epitopes defined by the amino acid sequences setforth in SEQ ID NOs: 88, 89, 90 and 91.

The antibody or its encoding cDNAs may be further modified. Thus, in afurther embodiment the method of the present invention comprises any oneof the step(s) of producing a chimeric antibody, single-chain antibody,Fab-fragment, bi-specific antibody, fusion antibody, labeled antibody oran analog of any one of those. Corresponding methods are known to theperson skilled in the art and are described, e.g., in Harlow and Lane“Antibodies, A Laboratory Manual”, CSH Press, Cold Spring Harbor, 1988.When derivatives of said antibodies are obtained by the phage displaytechnique, surface plasmon resonance as employed in the BIAcore systemcan be used to increase the efficiency of phage antibodies which bind tothe same epitope as that of any one of the antibodies described herein(Schier, Human Antibodies Hybridomas 7 (1996), 97-105; Malmborg, J.Immunol. Methods 183 (1995), 7-13). The production of chimericantibodies is described, for example, in international applicationWO89/09622. Methods for the production of humanized antibodies aredescribed in, e.g., European application EP-A1 0 239 400 andinternational application WO90/07861. As discussed above, the antibodyof the invention may exist in a variety of forms besides completeantibodies; including, for example, Fv, Fab and F(ab)₂, as well as insingle chains; see e.g. international application WO88/09344.

The antibodies of the present invention or their correspondingimmunoglobulin chain(s) can be further modified using conventionaltechniques known in the art, for example, by using amino aciddeletion(s), insertion(s), substitution(s), addition(s), and/orrecombination(s) and/or any other modification(s) known in the arteither alone or in combination. Methods for introducing suchmodifications in the DNA sequence underlying the amino acid sequence ofan immunoglobulin chain are well known to the person skilled in the art;see, e.g., Sambrook, Molecular Cloning A Laboratory Manual, Cold SpringHarbor Laboratory (1989) N.Y. and Ausubel, Current Protocols inMolecular Biology, Green Publishing Associates and Wiley Interscience,N.Y. (1994). Modifications of the antibody of the invention includechemical and/or enzymatic derivatizations at one or more constituentamino acids, including side chain modifications, backbone modifications,and N- and C-terminal modifications including acetylation,hydroxylation, methylation, amidation, and the attachment ofcarbohydrate or lipid moieties, cofactors, and the like. Likewise, thepresent invention encompasses the production of chimeric proteins whichcomprise the described antibody or some fragment thereof at the aminoterminus fused to heterologous molecule such as an immunostimulatoryligand at the carboxyl terminus; see, e.g., international applicationWO00/30680 for corresponding technical details.

A further aspect refers to a polynucleotide encoding the antibody orbinding fragment thereof as described herein. Another embodiment refersto a vector comprising a polynucleotide encoding the antibody or bindingfragment thereof as described herein. Another embodiment refers to acell comprising a polynucleotide encoding the antibody or bindingfragment thereof as described herein or a vector comprising apolynucleotide encoding the antibody or binding fragment thereof asdescribed.

In this respect, the person skilled in the art will readily appreciatethat the polynucleotides encoding at least the variable domain of thelight and/or heavy chain may encode the variable domains of bothimmunoglobulin chains or only one. Likewise, said polynucleotides may beunder the control of the same promoter or may be separately controlledfor expression. Possible regulatory elements permitting expression inprokaryotic host cells comprise, e.g., the P_(L), lac, trp or tacpromoter in E. coli, and examples for regulatory elements permittingexpression in eukaryotic host cells are the AOX1 or GAL1 promoter inyeast or the CMV-, SV40- , RSV-promoter, CMV-enhancer, SV40-enhancer ora globin intron in mammalian and other animal cells. Beside elementswhich are responsible for the initiation of transcription suchregulatory elements may also comprise transcription termination signals,such as the SV40-poly-A site or the tk-poly-A site, downstream of thepolynucleotide. Furthermore, depending on the expression system usedleader sequences capable of directing the polypeptide to a cellularcompartment or secreting it into the medium may be added to the codingsequence of the polynucleotide of the invention and are well known inthe art. The leader sequence(s) is (are) assembled in appropriate phasewith translation, initiation and termination sequences, and preferably,a leader sequence capable of directing secretion of translated protein,or a portion thereof, into the periplasmic space or extracellularmedium. Optionally, the heterologous sequence can encode a fusionprotein including a C- or N-terminal identification peptide impartingdesired characteristics, e.g., stabilization or simplified purificationof expressed recombinant product. Preferably, the expression controlsequences will be eukaryotic promoter systems in vectors capable oftransforming or transfecting eukaryotic host cells, but controlsequences for prokaryotic hosts may also be used. Once the vector hasbeen incorporated into the appropriate host, the host is maintainedunder conditions suitable for high level expression of the nucleotidesequences, and, as desired, the collection and purification of theimmunoglobulin light chains, heavy chains, light/heavy chain dimers orintact antibodies, binding fragments or other immunoglobulin forms mayfollow; see, Beychok, Cells of Immunoglobulin Synthesis, Academic Press,N.Y., (1979).

Furthermore, the present invention relates to vectors, particularlyplasmids, cosmids, viruses and bacteriophages used conventionally ingenetic engineering that comprise a polynucleotide encoding the antigenor preferably a variable domain of an immunoglobulin chain of anantibody of the invention; optionally in combination with apolynucleotide of the invention that encodes the variable domain of theother immunoglobulin chain of the antibody of the invention. Preferably,said vector is an expression vector and/or a gene transfer or targetingvector. Expression vectors derived from viruses such as retroviruses,vaccinia virus, adeno-associated virus, herpes viruses, or bovinepapilloma virus, may be used for delivery of the polynucleotides orvector of the invention into targeted cell population. Methods which arewell known to those skilled in the art can be used to constructrecombinant viral vectors; see, for example, the techniques described inSambrook, Molecular Cloning A Laboratory Manual, Cold Spring HarborLaboratory (1989) N.Y. and Ausubel, Current Protocols in MolecularBiology, Green Publishing Associates and Wiley Interscience, N.Y.(1994). Alternatively, the polynucleotides and vectors of the inventioncan be reconstituted into liposomes for delivery to target cells. Thevectors containing the polynucleotides of the invention (e.g., the heavyand/or light variable domain(s) of the immunoglobulin chains encodingsequences and expression control sequences) can be transferred into thehost cell by well-known methods, which vary depending on the type ofcellular host. For example, calcium chloride transfection is commonlyutilized for prokaryotic cells, whereas calcium phosphate treatment orelectroporation may be used for other cellular hosts; see Sambrook,supra.

The present invention furthermore relates to host cells transformed witha polynucleotide or vector of the invention. Said host cell may be aprokaryotic or eukaryotic cell. The polynucleotide or vector of theinvention which is present in the host cell may either be integratedinto the genome of the host cell or it may be maintainedextrachromosomally. The host cell can be any prokaryotic or eukaryoticcell, such as a bacterial, insect, fungal, plant, animal or human cell.Preferred fungal cells are, for example, those of the genusSaccharomyces, in particular those of the species S. cerevisiae. Theterm “prokaryotic” is meant to include all bacteria which can betransformed or transfected with a DNA or RNA molecules for theexpression of an antibody of the invention or the correspondingimmunoglobulin chains. Prokaryotic hosts may include gram negative aswell as gram positive bacteria such as, for example, E. coli, S.typhimurium, Serratia marcescens and Bacillus subtilis. The term“eukaryotic” is meant to include yeast, higher plant, insect andpreferably mammalian cells, most preferably HEK293, NSO and CHO cells.Depending upon the host employed in a recombinant production procedure,the antibodies or immunoglobulin chains encoded by the polynucleotide ofthe present invention may be glycosylated or may be non-glycosylated.Antibodies of the invention or the corresponding immunoglobulin chainsmay also include an initial methionine amino acid residue. Apolynucleotide of the invention can be used to transform or transfectthe host using any of the techniques commonly known to those of ordinaryskill in the art. Furthermore, methods for preparing fused, operablylinked genes and expressing them in, e.g., mammalian cells and bacteriaare well-known in the art (Sambrook, Molecular Cloning: A LaboratoryManual, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., 1989).The genetic constructs and methods described therein can be utilized forexpression of the antibody of the invention or the correspondingimmunoglobulin chains in eukaryotic or prokaryotic hosts. In general,expression vectors containing promoter sequences which facilitate theefficient transcription of the inserted polynucleotide are used inconnection with the host. The expression vector typically contains anorigin of replication, a promoter, and a terminator, as well as specificgenes which are capable of providing phenotypic selection of thetransformed cells. Suitable source cells for the DNA sequences and hostcells for immunoglobulin expression and secretion can be obtained from anumber of sources, such as the American Type Culture Collection(“Catalogue of Cell Lines and Hybridomas,” Fifth edition (1985)Rockville, Md., U.S.A., which is incorporated herein by reference).

Furthermore, transgenic animals, preferably mammals, comprising cells ofthe invention may be used for the large scale production of the antibodyof the invention.

A further aspect refers to a method for preparing an anti-allergenantibody or allergen-binding fragment thereof, consisting of culturingthe cell comprising a polynucleotide encoding the antibody or bindingfragment thereof as described herein and isolating the antibody orallergen binding fragment thereof from the cell or culture medium of thecell.

In a further embodiment, the present invention relates to a method forthe production of an antibody or a binding fragment thereof, said methodcomprising

(a) culturing a cell as described herein; and

(b) isolating said antibody or binding fragment thereof from theculture.

The transformed hosts can be grown in fermentors and cultured accordingto techniques known in the art to achieve optimal cell growth. Onceexpressed, the whole antibodies, their dimers, individual light andheavy chains, or other immunoglobulin forms of the present invention,can be purified according to standard procedures of the art, includingammonium sulfate precipitation, affinity columns, column chromatography,gel electrophoresis and the like; see, Scopes, “Protein Purification”,Springer Verlag, N.Y. (1982). The antibody or its correspondingimmunoglobulin chain(s) of the invention can then be isolated from thegrowth medium, cellular lysates, or cellular membrane fractions. Theisolation and purification of the, e.g., recombinantly expressedantibodies or immunoglobulin chains of the invention may be by anyconventional means such as, for example, preparative chromatographicseparations and immunological separations such as those involving theuse of monoclonal or polyclonal antibodies directed, e.g., against theconstant region of the antibody of the invention. It will be apparent tothose skilled in the art that the antibodies of the invention can befurther coupled to other moieties for, e.g., drug targeting and imagingapplications. Such coupling may be conducted chemically after expressionof the antibody or antigen to site of attachment or the coupling productmay be engineered into the antibody or antigen of the invention at theDNA level. The DNAs are then expressed in a suitable host system, andthe expressed proteins are collected and renatured, if necessary.

Substantially pure immunoglobulins of at least about 90 to 95%homogeneity are preferred, and 98 to 99% or more homogeneity mostpreferred, for pharmaceutical uses. Once purified, partially or tohomogeneity as desired, the antibodies may then be used therapeutically(including extracorporally) or in developing and performing assayprocedures.

The present invention also involves a method for producing cells capableof expressing an antibody of the invention or its correspondingimmunoglobulin chain(s) comprising genetically engineering cells withthe polynucleotide or with the vector of the invention.

Moreover, the invention relates to an antibody composition comprising atleast two antibodies, wherein at least one of the antibodies is selectedfrom the antibodies as defined herein. In one embodiment the antibodycomposition comprises at least two of the antibodies selected from theantibodies as defined herein. In one embodiment the antibody compositioncomprises at least three of the antibodies selected from the antibodiesas defined herein.

In one embodiment, the antibody composition comprises the antibodies32B10, 37D5 and 7G6.

Another aspect of the invention refers to a pharmaceutical compositioncomprising at least one of the compounds selected from the groupconsisting of antibody or binding fragment as described herein, or theantibody composition as described herein, the polynucleotide asdescribed herein, the vector as described herein and the cell asdescribed herein.

In one embodiment, the pharmaceutical comprises the antibody or bindingfragment of any as described herein or the antibody composition asdescribed herein.

In another embodiment the pharmaceutical composition further comprisesan additional agent useful for treating peanut allergy. The additionalagent useful for treating peanut allergy may be a β-adrenergic agonist,such as epinephrine, antihistamine, corticosteroid, anti-IgE antibody,anti-IgE antibody binding fragment, peptide vaccine and furtherantibodies capable of binding to a peanut allergen. Preferably, thepharmaceutical composition comprises epinephrine.

Such compositions pharmaceutical comprise a therapeutically orprophylactically effective amount of an antibody or binding fragmentthereof in admixture with a suitable pharmaceutical acceptable carrier,e.g., a pharmaceutically acceptable agent.

Pharmaceutically acceptable agents for use in the present pharmaceuticalcompositions include carriers, excipients, diluents, antioxidants,preservatives, coloring, flavoring and diluting agents, emulsifyingagents, suspending agents, solvents, fillers, bulking agents, buffers,delivery vehicles, tonicity agents, co-solvents, wetting agents,complexing agents, buffering agents, antimicrobials, and surfactants.

The composition can be in liquid form or in a lyophilized orfreeze-dried form and may include one or more lyoprotectants,excipients, surfactants, high molecular weight structural additivesand/or bulking agents (see for example U.S. Pat. Nos. 6,685,940,6,566,329, and 6,372,716).

Compositions can be suitable for parenteral administration. Exemplarycompositions are suitable for injection or infusion into an animal byany route available to the skilled worker, such as intraarticular,subcutaneous, intravenous, intramuscular, intraperitoneal, intracerebral(intraparenchymal), intracerebroventricular, intramuscular, intraocular,intraarterial, or intralesional routes. A parenteral formulationtypically will be a sterile, pyrogen-free, isotonic aqueous solution,optionally containing pharmaceutically acceptable preservatives.Preferably the pharmaceutical composition is intended for subcutaneous,intravenous, intramuscular, intraperitoneally intranasal and/orinhalative administration.

Examples of non-aqueous solvents are propylene glycol, polyethyleneglycol, vegetable oils such as olive oil, and injectable organic esterssuch as ethyl oleate. Aqueous carriers include water, alcoholic/aqueoussolutions, emulsions or suspensions, including saline and bufferedmedia. Parenteral vehicles include sodium chloride solution, Ringers'dextrose, dextrose and sodium chloride, lactated Ringer's, or fixedoils. Intravenous vehicles include fluid and nutrient replenishers,electrolyte replenishers, such as those based on Ringer's dextrose, andthe like. Preservatives and other additives may also be present, suchas, for example, anti-microbials, anti-oxidants, chelating agents, inertgases and the like. See generally, Remington's Pharmaceutical Science,16th Ed., Mack Eds., 1980, which is incorporated herein by reference.

Pharmaceutical compositions described herein can be formulated forcontrolled or sustained delivery in a manner that provides localconcentration of the product (e.g., bolus, depot effect) and/orincreased stability or half-life in a particular local environment. Thecompositions can include the formulation of antibodies, bindingfragments, nucleic acids, or vectors of the invention with particulatepreparations of polymeric compounds such as polylactic acid,polyglycolic acid, etc., as well as agents such as a biodegradablematrix, injectable microspheres, microcapsular particles, microcapsules,bioerodible particles beads, liposomes, and implantable delivery devicesthat provide for the controlled or sustained release of the active agentwhich then can be delivered as a depot injection.

Both biodegradable and non-biodegradable polymeric matrices can be usedto deliver compositions of the present invention, and such polymericmatrices may comprise natural or synthetic polymers. Biodegradablematrices are preferred. The period of time over which release occurs isbased on selection of the polymer. Typically, release over a periodranging from between a few hours and three to twelve months is mostdesirable.

Alternatively or additionally, the compositions can be administeredlocally via implantation into the affected area of a membrane, sponge,or other appropriate material on to which an antibody, binding fragment,nucleic acid, or vector of the invention has been absorbed orencapsulated. Where an implantation device is used, the device can beimplanted into any suitable tissue or organ, and delivery of anantibody, binding fragment, nucleic acid, or vector of the invention canbe directly through the device via bolus, or via continuousadministration, or via catheter using continuous infusion.

A pharmaceutical composition comprising a binding antibody or bindingfragment thereof can be formulated for inhalation, such as for example,as a dry powder. Inhalation solutions also can be formulated in aliquefied propellant for aerosol delivery. In yet another formulation,solutions may be nebulized.

Certain formulations containing antibodies or binding fragments thereofcan be administered orally. Formulations administered in this fashioncan be formulated with or without those carriers customarily used in thecompounding of solid dosage forms such as tablets and capsules. Forexample, a capsule can be designed to release the active portion of theformulation at the point in the gastrointestinal tract whenbioavailability is maximized and pre-systemic degradation is minimized.Additional agents can be included to facilitate absorption of aselective binding agent. Diluents, flavorings, low melting point waxes,vegetable oils, lubricants, suspending agents, tablet disintegratingagents, and binders also can be employed.

The phrase “therapeutically or prophylactically effective amount” asused herein refers to an amount that produces the desired effect forwhich it is administered. The exact amount will depend on the purpose ofthe treatment, and will be ascertainable by one skilled in the art usingknown techniques (see, for example, Lloyd (2012) The Art, Science andTechnology of Pharmaceutical Compounding).

Other aspects of the invention refer to the antibodies or bindingfragments, antibody composition, polynucleotide, vector, cell orpharmaceutical composition as described herein for use in the treatmentof allergy. The allergy may be a food allergy, such as peanut allergy.In particular, the invention refers to the antibodies or bindingfragments as described herein, the antibody composition as describedherein and the pharmaceutical composition as described herein for use inthe treatment of food allergy, such as peanut allergy. The treatment maybe prophylactic or therapeutic.

Correspondingly, the application relates to methods of treatingallergies, in particular food allergies, such as peanut allergy byadministering the antibodies or binding fragments, antibody composition,polynucleotide, vector, cell or pharmaceutical composition according tothe invention. Preferably, antibodies or binding fragments thereof, theantibody composition and the pharmaceutical composition according to theinvention are administered.

Further, the application relates to an antibody or binding fragmentthereof, antibody composition, polynucleotide, vector, cell orpharmaceutical composition according to the invention in the manufactureof a medicament for the treatment of allergies, in particular foodallergies, such as peanut allergy.

Another aspect refers to a method of evaluating the capacity of ancandidate antibody or binding fragment thereof to inhibit allergenbinding/and/or allergen-induced activity in an human, wherein the methodcomprises

(i) incubating the candidate antibody or binding fragment thereof, acomposition comprising IgEs derived from the human and a food allergen;

(ii) evaluating whether the candidate antibody inhibits allergen bindingto IgEs/and/or allergen-induced activity in the composition comprisingIgEs derived from the human.

The term “evaluating” comprises “detecting” or “determining” which leadyes or no output and to “measuring” which gives numerical values, suchas continuous data values, as output.

In one embodiment the method further comprises the step of

(iii) determining whether the administration of the candidate antibodyis a suitable treatment for a patient suffering from food allergy basedon the result of step (ii).

The administration of the candidate antibody is considered as suitabletreatment for a patient suffering from food allergy, if the candidateantibody inhibits allergen binding to IgEs and/or inhibitsallergen-induced activity in the composition comprising IgEs derivedfrom the human.

In other words, the invention refers to an in vitro method fordetermining efficacy of treatment of a subject suffering from foodallergy, in particular peanut allergy by treatment with an antibody orfragment thereof according to the invention, comprising determining invitro whether the candidate antibody inhibits allergen binding to theIgEs and/or inhibits allergen-induced activity in the compositioncomprising IgEs derived from the human.

Therefore, the present application refers to methods for determiningwhether a patient suffering from food allergy is responsive to acandidate antibody, comprising the steps of

(i) incubating the candidate antibody or binding fragment thereof, acomposition comprising IgEs derived from the patient and an allergen;

(ii) evaluating whether the candidate antibody inhibits allergenbinding/and/or allergen-induced activity in the composition comprisingIgEs derived from the patient;

(iii) determining whether the administration of the candidate antibodyis a suitable treatment for the patient suffering from food allergybased on the result of step (ii).

In a specific embodiment, the present application refers to a method fordetermining whether a patient suffering from food allergy is responsiveto a candidate antibody, comprising the steps of

(i) incubating the candidate antibody or binding fragment thereof, acomposition comprising IgEs derived from the patient and an allergen;

(ii) evaluating whether the candidate antibody inhibits allergen bindingand/or allergen-induced activity in the composition comprising IgEsderived from the patient by measuring the level of activated leukocytes;

(iii) determining whether the administration of the candidate antibodyis a suitable treatment for the patient suffering from food allergybased on the result of step (ii).

wherein decreased levels of activated leukocytes compared to a controlindicate that the antibody is capable of inhibiting allergen bindingand/or allergen induced activity in the human patient;

Thus, the invention refers to the antibody or binding fragment thereofaccording to the invention for use in the treatment of food allergy, inparticular peanut allergy, of a human wherein the antibody isadministered if the antibody or binding fragment thereof inhibitsallergen binding to the IgEs and/or inhibits allergen-induced activityin the composition comprising IgEs derived from the human. The skilledperson understands that the test whether or not the antibody or bindingfragment thereof inhibits allergen binding to the IgEs and/or inhibitsallergen-induced activity in the composition comprising IgEs derivedfrom the human is carried out in vitro.

The skilled person is aware of methods for testing the capability of theantibody or binding fragment thereof to compete for the allergen (asdescribed for example in Uermosi et al., Mechanism of allergen-specificdesensitization, Allergy, 2010) For example a competition ELISA assay asdescribed in example 4 could be used. In brief, allergen coated platesare incubated with increasing concentration of the antibodies to betested, e.g. for 1 h at room temperature. Subsequently plates areincubated with the composition comprising the IgEs derived from thehuman. Total IgE levels can be detected using an appropriate antibodythat specifically binds to IgE.

Thus, one embodiment refers to a method of evaluating the capability ofa candidate antibody or binding fragment thereof to inhibit allergenbinding, comprising the following steps:

(i) incubating the candidate antibody or binding fragment thereoftogether with a composition comprising IgEs derived from the human and afood allergen;

(ii) measuring the level of IgE bound to antigen;

wherein decreased levels of IgE bound to antigen compared to a controlindicate that the antibody is capable of inhibiting allergen binding.

Thereby, decreased levels of IgE bound to antigen compared to thecontrol indicated that the antibody is suitable for the treatment of apatient suffering from food allergy.

Typically, the control comprises IgEs derived from the human and a foodallergen but does not contain the candidate antibody. The candidateantibody is an antibody or binding fragment thereof as defined herein.

The skilled person is aware of methods for testing the capability of theantibody or binding fragment thereof to bind to the allergen (seeexample 2). For example a binding ELISA assay could be used. In brief,allergen coated plates are incubated with increasing concentration ofthe antibodies to be tested, e.g. for 1 h at room temperature. Anappropriate detection antibody is used to measure concentrations/levelsof anti-allergen antibody.

The determination whether the antibodies or fragments thereof of theinvention inhibit allergen-induced activity in the compositioncomprising IgEs derived from the human will be ascertainable by oneskilled in the art using known techniques (see, for example Hausmann etal., Robust expression of CCR3 as single basophil marker', Allergy,2011) .

For example the Basophil activation test could be used. In brief theallergen was preincubated with one or serial dilutions of the candidateantibody. Afterwards, 100 μl of whole blood of the patient werestimulated with the allergen or stimulation buffer (negative control) inthe presence/absence of the candidate antibody or isotype control for 30minutes at 37° C., 5% CO2. Cells are simultaneously stained withanti-CCR3-APC, anti-CD203c-PE and anti-CD63-FITC (Biolegend, San Diego,Calif., USA). Basophils are gated as SSClow, CCR3high lymphocytes. Atleast 500 basophils are acquired using e.g. a FACSCalibur (BectonDickinson AG, Allschwil, Switzerland). Activation of basophils isquantified using % of CD63+ CCR3+ basophils.

The term “isotype control” as used herein refers to an antibody of thesame isotype and having the same constant region as the candidateantibody but being not specific for the allergen.

Thus, one embodiment refers to a method of evaluating the capability ofa candidate antibody or binding fragment thereof to inhibit allergenbinding and/or allergen-induced activity in a human, comprising thefollowing steps:

(i) incubating the candidate antibody or binding fragment thereoftogether with basophils derived from the human and a food allergen;

(ii) measuring the level of activated basophils;

wherein decreased levels of activated basophils compared to a controlindicate that the antibody is capable of inhibiting allergen bindingand/or allergen induced activity in a human.

Thereby, decreased levels of activated basophils compared to the controlindicated that the antibody is suitable for the treatment of a patientsuffering from food allergy.

Typically, the control comprises basophils derived from the human and afood allergen but does not contain the candidate antibody.

Preferably the candidate antibody is an antibody or binding fragmentthereof according to the invention.

The skilled person understands that the step of determining whethercandidate antibody inhibits allergen binding to the IgEs and/or inhibitsallergen-induced activity in the composition comprising IgEs alsoencompasses that several candidate antibodies or fragments thereof aretested on the same sample together (are pooled) in a first step, and ifthe result is positive (i.e. that the allergen binding and/orallergen-induced activity is inhibited), then in a second step thecandidate antibodies or fragments thereof are tested individually.Alternatively, the candidate antibodies are already tested individuallyin the first step.

Preferably, in step (i) the composition comprising IgEs from the human,in particular the allergic patient comprises basophils. Typically, thebasophils are derived from the allergicpatient. Alternatively, thebasophils are donor-derived IgE-stripped basophils.

IgE-stripped basophils” may be produced by incubating basophils withlactic acid, in order to strip off the IgEs from the donor. In order tofurnish the stripped basophils with patient derived IgEs, theIgE-stripped basophils may be incubated with plasma from an allergicpatient.

For the identification of basophils, in particular activated basophils,cell sorting such as fluorescent activated cell sorting could be used.In one embodiment, basophils are identified based on the lowside-scattered light (SSC) and the expression of CCR3. In addition,activated basophils are identified by measuring surface expression ofCD63 and/or surface expression of CD203c. Typically, in activatedbasophils the surface expression of CD63 and/or CD203c is high.

The composition comprising IgEs derived from the human may be plasma,sera, blood, saliva, peripheral blood mononuclear cell (PBMC),leukocytes, basophils or IgEs stripped from basophils.

In a specific embodiment, evaluating the capability of a candidateantibody or binding fragment thereof to inhibit allergen binding and/orallergen-induced activity is accomplished by an assay measuring aninflammatory mediator released from the basophil, preferablyleukotriene. This assay is advantageous, since the measurement of theleukotriene released from the basophils can be detected by ELISA andtherefore can be easily carried out in high-throughput format. Theleukotriene release assay allows to evaluate the capability of acandidate antibody or binding fragment thereof to inhibit allergenbinding and/or allergen-induced activity with high sensitivity,reliability and cost effectiveness.

In assays which determine the basophil activation by measuring thesecretion of the inflammatory mediator, in particular leukotriene, fromthe basophils, the IgE containing composition may be isolatedleukocytes. The leukocytes may be either derived from an allergicpatient or may be donor-derived leukocytes, from which the IgEs from thedonor are stripped-off and replaced by IgEs derived from an allergicpatient.

The skilled person understands that leukotrienes may be secreted also bymast cells, eosinophils, neutrophils. However, in this experimentalset-up, the leukotriene secretion from these cell types is neglectablein comparison to the secretion of leukotrienes by basophils.

The leukotriene assay is described in detail in the examples. In brief,leukocytes may be isolated by dextran sedimentation from whole blood ofallergic patients. Titrations of peanut extract are preincubated withpurified antibody samples. Afterwards, leukocytes are stimulated withtitrated peanut extract or stimulation buffer (negative control) in thepresence/absence of different anti-peanut allergen antibodies or isotypecontrol. Activation of leukocytes, in particular, basophils isquantified using CAST®ELISA (Balmann, Schonenbuch, Switzerland).

“Leukotriene” as used herein refers to a family of eicosanoidinflammatory mediators. In particular, the term “leukotriene” refers tosulfidoleukotrienes, also termed cysteinyl leukotrienes, such as LTC₄and its metabolites LTD₄ and LTE₄.

Leukotriene is released by activated basophils. Thus decreasedleukotriene levels detected for the candidate antibody or bindingfragment thereof compared to a control sample indicate that the antibodyis capable of inhibiting allergen binding and/or allergen inducedactivity in a human.

Leukotriene is released from the basophils in a dose dependent manner.Thus, it is particularly suitable for highly sensitive assays.

Therefore, the methods described above can be used to compare thedifferent candidate antibodies for the capability to treat peanutallergy, wherein a first candidate antibody leading to a loweractivation of the basophils, e.g. indicated by a lower secretion ofleukotriene in comparison to a second candidate antibody, indicates thatthe first candidate antibody is particularly suited for treating peanutallergy.

Accordingly, one embodiment refers to a method of evaluating thecapability of a candidate antibody or binding fragment thereof toinhibit allergen binding and/or allergen-induced activity in a human,comprising the following steps:

(i) incubating the candidate antibody or binding fragment thereoftogether with a composition comprising IgEs derived from the human and afood allergen;

(ii) evaluating whether the candidate antibody inhibits allergenbinding/and/or allergen-induced activity in the composition comprisingIgEs derived from the patient by measuring the leukotriene secretion;

wherein decreased levels of leukotriene secretion compared to a controlindicate that the antibody is capable of inhibiting allergen bindingand/or allergen induced activity in a human.

In another preferred embodiment, the present application refers to amethod for determining whether a patient suffering from food allergy isresponsive to a candidate antibody, comprising the steps of

(i) incubating the candidate antibody or binding fragment thereof, acomposition comprising IgEs derived from the patient and an allergen;

(ii) evaluating whether the candidate antibody inhibits allergenbinding/and/or allergen-induced activity in the composition comprisingIgEs derived from the patient by measuring the leukotriene secretion;

(iii) determining whether the administration of the candidate antibodyis a suitable treatment for the patient suffering from food allergybased on the result of step (ii).

wherein decreased levels of leukotriene secretion compared to a controlindicate that the antibody is capable of inhibiting allergen bindingand/or allergen induced activity in the human patient;

Another aspect of the invention refers to a method of detecting orquantifying whether an allergen is present in a sample comprising thefollowing steps:

i) incubation of the sample with an antibody of the invention asdescribed herein or with an antibody composition of the invention asdescribed herein,

ii) detecting the antibody which is bound to allergen in the sample.

The antibody may be detectably labeled. Alternatively, the antibody isused with a second antibody that is detectably labeled. Preferably, theantibody is unlabeled and used in combination with a second antibodythat is detectably labeled. The detectable label may be selected fromthe group consisting of an enzyme, a radioisotope, a fluorophore, apeptide and a heavy metal.

Labeling agents can be coupled either directly or indirectly to theantibodies or antigens of the invention. One example of indirectcoupling is by use of a spacer moiety. Furthermore, the antibodies ofthe present invention can comprise a further domain, said domain beinglinked by covalent or non-covalent bonds. The linkage can be based ongenetic fusion according to the methods known in the art and describedabove or can be performed by, e.g., chemical cross-linking as describedin, e.g., international application WO94/04686. The additional domainpresent in the fusion protein comprising the antibody of the inventionmay preferably be linked by a flexible linker, advantageously apolypeptide linker, wherein said polypeptide linker comprises plural,hydrophilic, peptide-bonded amino acids of a length sufficient to spanthe distance between the C-terminal end of said further domain and theN-terminal end of the antibody of the invention or vice versa. Thetherapeutically or diagnostically active agent can be coupled to theantibody of the invention or an antigen-binding fragment thereof byvarious means. This includes, for example, single-chain fusion proteinscomprising the variable regions of the antibody of the invention coupledby covalent methods, such as peptide linkages, to the therapeutically ordiagnostically active agent. Further examples include molecules whichcomprise at least an antigen-binding fragment coupled to additionalmolecules covalently or non-covalently include those in the followingnon-limiting illustrative list.

Experiments

EXAMPLES

The following examples further illustrate the invention, but should notbe construed to limit the scope of the invention in any way. Detaileddescriptions of conventional methods, such as those employed herein canbe found in the cited literature. The practice of the present inventionwill employ, unless otherwise indicated, conventional techniques of cellbiology, cell culture, molecular biology, transgenic biology,microbiology, recombinant DNA, and immunology, which are within theskill of the art.

Material and Methods

Patients Selection and Disease Associations of the Invention

As starting material for the cloning of fully human antibodies, humanlymphocytes were obtained from peripheral blood of 84 voluntary allergicpatients . All patients gave their written informed consent and thestudy has been approved by the ethical Committee of Zurich. Clinicalreactivity was defined based on anamnesis of signs of allergic reactions(e.g. oral allergy syndrome, angioedema, dyspnea, nausea, emesis,diarrhea, shock) and/or positive skin prick test or presence of serumIgE antibodies (ImmunoCAP, Phadia, Uppsala, Sweden).

Isolation of Memory B Cells Reactive to Peanut Allergens

Antibodies specific to major peanut allergens were isolated by molecularcloning of immunoglobulin genes obtained from single-cell sorted cellsderived from short term oligoclonal cultures of activated memory B cellsproducing the antibodies of interest.

Isolation of PBMC and memory B cell culture

Peripheral blood mononuclear cells (PBMCs) were isolated fromheparinized peripheral blood using Lympholyte H according tomanufacturer's instruction (Cedarlane, Burlington, Ontario, Canada) andcryopreserved prior to use.

Cells were stained on ice with monoclonal antibodiesphycoerythrin-conjugated anti-human IgD and IgA, APC-conjugated mAbsanti-human IgM, CD3, CD56, CD8, and FITC-conjugated mAb anti-human CD22(Becton Dickinson, Basel, Switzerland). Cells sorting was carried outusing a MoFlo XDP cell sorter (Beckman Coulter, Krefeld, Germany). CD22positive and IgM, IgD, IgA negative B cells were seeded at 5-10 cellsper well on irradiated CD40L-expressing feeder cells stimulated with acytokine cocktail, as described in Huang et al., Isolation of humanmonoclonal antibodies from peripheral blood cells', Nature Protocols,2012.

After 10-14 days of stimulation, culture supernatants were screened forthe presence of IgG or IgE antibodies specific for the target ofinterest (e.g. Ara h 1, Ara h 2, Ara h 3, Ara h 6, peanut extract). Thescreening process comprised for binding on recombinant or naturallyextracted peanut allergens of the particular molecule of interest (e.g.by ELISA). Detection of peanut-specific IgE or IgG antibodies isperformed using anti-human HRP-conjugated Fc-epsilon-specific secondaryantibody (Abcam, Cambridge, UK) or anti-human HRP-conjugated goatFc-gamma-specific antibody (Jackson ImmunoResearch, West Grove, Pa.,USA) followed by measurement of the HRP activity using atetramethylbenzidine substrate solution (TMB, Sigma-Aldrich Chemie GmbH,Buchs, Switzerland). Subsequently the antibody for which binding isdetected or the cell producing said antibody is isolated.

Molecular Cloning of Human Antibodies Specific to Peanut Allergens

Molecular cloning of human antibodies specific to peanut allergens iscarried out according to Huang et al., Isolation of human monoclonalantibodies from peripheral blood cells', Nature Protocols, 2012. Inparticular, single cells obtained from peanut allergen-reactive memory Bcell cultures are sorted into a 96 well PCR plate, containingreverse-transcription buffer (Invitrogen, Carlsbad, Calif., UnitedStates). cDNA preparation is done using Random Hexamer Primer(Invitrogen, Carlsbad, Calif., United States) according to thesupplier's protocol. Immunoglobulin heavy and light chain variableregions (V_(II) and V_(L), respectively) are PCR amplified according tostandard protocols (Wardemann et al, Science 301, 2003, 1374-1377). Toincrease the PCR efficiency, a nested PCR is performed. The first roundPCR is performed with primers specific for the IgG and IgE constantregion and primer mixes specific to all leader sequences from V_(H) andV_(L) families (Wardemann et al, Science 301, 2003, 1374-1377).Subsequently, a nested PCR is performed using primer mixes specific tothe 5′ region of framework 1 of V_(H) and V_(L) families (V-region) andthe immunoglobulin J-regions. Sequence analysis is carried out toidentify individual antibody clones present in the selected B cellculture. Afterwards, the V_(H) and V_(L) of each unique antibody cloneare cloned into expression vectors providing the constant regions ofhuman IgG 1 , human Ig-Kappa or human Ig-Lambda. Upon co-transfection ofthe Ig-heavy- and light expression vectors into HEK293T cells, therespective antibody clones are produced. Identification of the antibodyclone presumably responsible for the anti-peanut reactivity of theparental B cell culture is performed upon re-screening of therecombinant antibodies for reactivity to peanut allergens in an ELISA.In order to identify and to correct primer encoded sequence mismatchesin the Ig-variable regions, an additional PCR amplification on originalcDNA of the reactive clone is performed using a semi-nested PCR.Therefore, primer mixes specific for the Ig heavy and light chain leadersequences (5′-primers) and two primer pairs specific for a conservedregion of the Ig heavy and light chain constant regions (3′-primers) areused. PCR products are cloned into pCRT™ 2.1-TOPO® vector (Invitrogen,Carlsbad, Calif., United States) and subsequently sequenced usingstandard primers. Alternatively, PCR products are directly subjected tosequencing using internal primers specific to conserved regions of theconstant domains of heavy and light chains. Sequence determination andannotation of the complete antibody is carried out and this informationis used to design specific primers for the cloning of the authentichuman V_(H) and V_(L) sequence into antibody (IgG) expression vectors.This approach also allows the identification of Ig isotype (andsubclass) of each isolated, reactive monoclonal antibody. These V_(H)and V_(L) sequences are then used for the production of recombinantantibodies which are subsequently characterized in more detail.

The sequences of the antibodies are set out in Table 1.

TABLE 1Antibody sequences, Underlined, bold nucleotides or amino acids indicate the CDRsin the sequence of the variable domain. Italic nucleotides or amino acids indicate sequences which have not been sequenced but obtained from database.Nucleotide and amino acid sequences of variable heavy (VH) andvariable light (VL), constant heavy (CH) and constant light  Seq.Antibody (CL) chains. ID 37D5 VHgaggaacagctggtggagtctgggggacgtttattacggcctggggagtccctgagcctctcctgt 12DNA gcagccgttggattcaacttcggt gattttggcatggcc tgggtccgccaacctccagggaaggggctggagtgggtcgct ggcatcaattggagtggccatagtacaggttttgtagactccatgaa gggtcgactcaccatctccagagacagcgccaagagttccctgtttctgcaaatgaacagtctgcgaggcgaggacacggccgtgtattactgtgcgaga gtcgggagactttgtagtggaatatttgcgactcaatgggtgctttgatctg tggggccaggggacaatggtcaccgtctcttca 37D5 VHEEQLVESGGRLLRPGESLSLSCAAVGFNFG DFGMA WVRQPPG  8 amino acid KGLEWVAGINWSGHSTGFVDSMKG RLTISRDSAKSSLFLQM NSLRGEDTAVYYCAR VGRLCSGDICDSMGAFDLWGQGTMV TVSS 37D5 VLgacatccagatgacccagtctccttccaccctgtctgcatctataggagacagagtcaccatcactt 11kappa-type gt cgggccagtcagaccattgacaactggttggcctggtatcaacagagaccagggaaagcc DNA cctaaactcctgatctat caggcgtctagtctacaaagtggggtctcatcaaggttcagaggcagtggatctggcacagaattcactctcaccatcaccagcctgcagcctgatgactttgctacttattattgtcagaagtctaatggctattctcgtact ttcggccaggggaccaaggtggagatcaaa 37D5 VLDIQMTQSPSTLSASIGDRVTITC RASQTIDNWLA WYQQRPGK  7 kappa-type APKLLIYQASSLQS GVSSRFRGSGSGTEFTLTITSLQPDDFATY amino acid YC QKSNGYSRTFGQGTKVEIK 37D5 CHgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcac  14IgG1 agcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcasubclassggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctc DNAagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagaaagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggatgagctgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaatga 37D5 CHASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS 10 IgG1GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN subclassHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPK amino acidPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 37D5 CLcgaactgtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgc 13kappa-typectctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataac DNAgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgttag 37D5 CLRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK  9 kappa-typeVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV amino acidYACEVTHQGLSSPVTKSFNRGEC 32B10 VHcaggtgcagctggtgcagtctggggctgaggtgaagaagcctggggcctcagtgaaggtctcct 26 DNAgcagggcttctggatacatcttccgc aactttgatatcaact gggtgcgacaggccactggacaagggcttgagtgga tgggatgagccctaagagtggtgacaccggctatgctcagaagtt ccagggcagggtcaccatgaccagggacacctccataaacacagcctacatggaactgagcagcctgacatctgaggattcggccgtctattactgtgcgaga ggtgtcgacgggaccaac tggggccagggaacccgggtcaccgtctcctca 32B10 VH QVQLVQSGAEVKKPGASVKVSCRASGYIFRNFDIN WVRQAT 22 amino acid GQGLEWMG WMSPKSGDTGYAQKFQG RVTMTRDTSINTAYMELSSLTSEDSAVYYCAR GVDGTN WGQGTRVTVSS 32B10 VLgacatcgtgatgacccagtctccagactccctgggtgtgtctctgggcgagagggccaccatcag  25kappa-type ctgc aagtccagccagagtattttagataactccaacaataagaacttcatagctggggttcca DNA gcagaaaccaggacagccccctaagctgctcatttactgggcatctgcccgggaatcc ggggtccctgaccgattcagtggcagcgggtctgggacagaattcactctcaccatcaacagcctgcaggctgaagatgtggcagtttattactgt taccaatactattctactcctcacact tttggccaggggaccaagctggatctcaga 32B10 VL DIVMTQSPDSLGVSLGERATISC KSSQSILDNSNNKNFIA WFQ21 kappa-type QKPGQPPKLLIY WASARES GVPDRFSGSGSGTEFTLTINSLQA amino acidEDVAVYYC YQYYSTPHT FGQGTKLDLR 32B10 CHgcttccaccaagggcccatccgtcttccccctggcgccctgctccaggagcacctccgagagcac  28IgG2 agccgccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcagsubclassgcgctctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctca DNAgcagcgtggtgaccgtgacctccagcaacttcggcacccagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggacaagacagttgagcgcaaatgttgtgtcgagtgcccaccgtgcccagcaccacctgtggcaggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacgtgcgtggtggtggacgtgagccacgaagaccccgaggtccagttcaactggtacgtggacggcgtggaggtgcataatgcaaagacaaagccacgggaggagcagttcaacagcacgttccgtgtggtcagcgtcctcaccgttgtgcaccaggactggctgaacggcaaggagtacaagtgcaaggtctccaacaaaggcctcccagcccccatcgagaaaaccatctccaaaaccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctaccccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaaggccacacctcccatgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaatga 32B10 CHASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNS 24 IgG2GALTSGVHTFPAVLQSSGLYSLSSVVTVTSSNFGTQTYTCNVD subclassHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDT amino acidLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKATPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSC SVMHEALHNHYTQKSLSLSPGK32B10 CLcgaactgtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgc 27kappa-typectctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataac DNAgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgttag 32B10 CLRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK 23 kappa-typeVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV amino acidYACEVTHQGLSSPVTKSFNRGEC 2F8 VHgaagatgtccagtgtgaggtgcagctggtggagtctgggggaggcttggtcaagccggggggg 40 DNAtcgctgagactctcctgtgcagcgtctggattcatcttcagc gattataacatgaa ttgggtccgccaggctccagggaaggggctggagtgggtttca tccattactagaagtagtaggaccatttactacgcagactctgtgaagggc cgattcaccatatccagagacaatgccaagaactcactgcatctgcaaatgaacagtctcagagacgcggacacggctgtgtattattgtgcgaga gaggatttcgatgtttcgactggcccctactacatggacgtc tggggcaacgggaccacggtcatcgtctcctca 2F8 VHEVQLVESGGGLVKPGGSLRLSCAASGFIFS DYNMN WVRQAP 36 amino acid GKGLEWVSSITRSSRTIYYADSVKG RFTISRDNAKNSLHLQM NSLRDADTAVYYCAR EDFDVSTGPYYMDVWGNGTTVIVSS 2F8 VLgaaattgtgttgacgcagtctccaggcaccctgtctttgtctccaggggaaagagccaccctctcct  39kappa-type gc aggggccagtcagagtgttagcaacatgttcttagtcggtatcagcagaaacctggccagg DNA ctcccaggctcctcatgtat ggtgcatctaccagggccactgacatcccagacaggttcagtggcagtgggtctgggacagacttcactctcaccatcagcagactggagcctgaagattttgcagtgtattactgt cagcagaatggtaactcaccatacac ttttggccaggggaccaagctggagatcaaa 2F8 VLEIVLTQSPGTLSLSPGERATLSC RASQSVSNMFLV WYQQKPG 35 kappa-type QAPRLLMYGASTRAT DIPDRFSGSGSGTDFTLTISRLEPEDFA amino acid VYYC QQNGNSPYTFGQGTKLEIK 2F8 CHgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcac 42IgG1 agcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcasubclassggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctc DNAagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatccegggaggagatgaccaagaaccaggtcagectgacctgcctggtcaaaggettctateccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctatagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtccccgggtaaatga 2F8 CHASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS 38 IgG1GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN subclassHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPK amino acidPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 2F8 CLcgaactgtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgc 41kappa-typectctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataac DNAgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgttag 2F8 CLRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK 37 kappa-typeVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV amino acidYACEVTHQGLSSPVTKSFNRGEC 7G6 VHcaggtgcagctggtggagtctgggggaggcgtggtgcagcctgggaggtccctgagactctcat 54 DNAgtgcagcctctggcatcgccttcaat gactacactatgcac tgggtccgccggtctccagacaagggcctggagtgggtggca gctatatcatatggtgggactaataaatactacgcagattccgtg aagggccgattcaccatetccagagacagttccaagaacaccctgtttctgcagatggacagcctgagagttgaggacacggctgtgtattactgtgcgaga gattctggttatcggagtcttttgcac tggggccagggaaccctggtcaccgtctcctca 7G6 VH QVQLVESGGGVVQPGRSLRLSCAASGIAFNDYTMH WVRRSP 50 amino acid DKGLEWVA AISYGGTNKYYADSVKG RFTISRDSSKNTLFLQMDSLRVEDTAVYYCAR DSGYRSLLH WGGTLVTVSS 7G6 VLgagattgtgttgactcagtctccactctccctgcccgtcacccctggtgagccggcctccatctcctg 53kappa-type c aggtcgagtcagagcctcgtgcatagaaatggatacaactatttagattggtacctgcagaa DNA gccagggcagtctccacagctcctgatctatatatggcttctaaacgggcctcc ggggtccctgacaggttcagtggcagtgggtcaggcacagaatttacactgaaaatcagcagagtggaggctgaggatgttggaatttattactgc atgcaagctctacaaactggacg tteggccaagggaccaaggtggaagtcaac 7G6 VL EIVLTQSPLSLPVTPGEPASISC RSSQSLVHRNGYNYLD WYLQ 49kappa-type KPGQSPQLLIY MASKRAS GVPDRFSGSGSGTEFTLKISRVEAE amino acidDVGIYYC MQALQTWT FGQGTKVEVN 7G6 CHgcttccaccaagggcccatccgtcttccccctggcgccctgctccaggagcacctccgagagcac  56IgG4 agccgccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcagsubclassgcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctca DNAgcagcgtggtgaccgtgccctccagcagcttgggcacgaagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggacaagagagttgagtccaaatatggtcccccatgcccatcatgcccagcacctgagttcctggggggaccatcagtcttcctgttccccccaaaacccaaggacactctcatgatctcccggacccctgaggtcacgtgcgtggtggtggacgtgagccaggaagaccccgaggtccagttcaactggtacgtggatggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagttcaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaacggcaaggagtacaagtgcaaggtctccaacaaaggcctcccgtcctccatcgagaaaaccatctccaaagccaaagggcagccccgagagccacaggtgtacaccctgcccccatcccaggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctaccccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctecttettectetacagcaggctaaccgtggacaagagcaggtggcaggaggggaatgtettetcatgetccgtgatgcatgaggetctgcacaaccactacacacagaagagcctctecctgtctctgggtaaatga 7G6 CH ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNS 52IgG4 GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVD subclassHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKD amino acidTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK 7G6 CLcgaactgtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgc 55kappa-typectctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataac DNAgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagettcaacaggggagagtgttag 7G6 CLRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK 51 kappa-typeVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV amino acidYACEVTHQGLSSPVTKSFNRGEC 4B2 VHcaggtgcagctgcaggaggegggcccacgactggtgaagccttcacagaccctgtcagtcacc 68 DNAtgcactgtctctggtgcctccatcacc attggcggttactactggagt tggatccgccagcacccagggaagggcctggaatggatgggg tacatctattccaatgggauacctactacaatccgtccctcaagagt cgaattgccatgtcaatagacacgtetaaaaaccagttctecctgaagetgacttctgtgacagccgcggacacggccatatatttctgtgcgcgg gaggcgtgggagacgccactg tggggccagggaaccctgatcaccgtctcctcc 4B2 VH QVQLQEAGPRLVKPSQTLSVTCTVSGASITIGGYYWS WIRQHP 64 amino acid GKGLEWMG YIYSNGRTYYNPSLKSRIAMSIDTSKNQFSLKLT SVTAADTAIYFCAR EAWETPL WGQGTLITVSS 4B2 VLgacatccagatgactcagtctccatcctccctgtctgcttctgtaggagacagagtcaccatcacttg 67kappa-type c caggcgaatcaggacattgtcaactctttaaattggtttcaacacaaaccagggacagcccct DNA aaagtcctgatctac gatgcatccaaattggaaacaggggtcccatctaggttcagtggaagtgggtctgggacacattttactttcaccataagtggcctgcagcctgaagattttgcaacatatttctgtcaacaatatgagaatcttccgcacacttttggccaggggaccaagttggagatcaga 4B2 VLDIQMTQSPSSLSASVGDRVTITC QANQDIVNSLN WFQHKPGT 63 kappa-type APKVLIYDASKLET GVPSRFSGSGSGTHFTFTISGLQPEDFATY amino acid FC QQYENLPHTFGQGTKLEIR 4B2 CHgcctccaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctgggggcac 70IgG1 agcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcasubclassggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctcaggactctactccctc DNAagcagcgtggtgaccgtgccctccagcagcttgggcacccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagaaagttgagcccaaatettgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacatgcgtggtggtggacgtgagccacgaagaccctgaggtcaagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggatgagctgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcctcccgtgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaatga 4B2 CHASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS 66 IgG1GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN subclassHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPK amino acidPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGK4B2 CLcgaactgtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgc 65kappa-typectctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtggataac DNAgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgttag 4B2 CLRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK 65 kappa-typeVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV amino acidYACEVTHQGLSSPVTKSFNRGEC 12G10 VHgaggtgcagctgttggagtctgggggacgtttattacggccgggggggtccctgagcctctcctg  80DNA tgtagcctctggattctacttcggt gattttggcatgagc tgggtccgccaggttccagggaaggggctggagtgggtctct ggcattgactggagtggccgtagtacaggttatgtagactccatgaa gggccgactcaccatctccagagacaacgacaagagttccctgtatttgcaaatgaacgatctgcgcggcgaggacacggccgtctattactgtgcgagg gtcgggagactctgtagtggtgattcttgc  gactcaatgggcgcttttgacctg tggggccaggggacaatggtcaccgtctcttca 12G10 VHEVQLLESGGRLLRPGGSLSLSCVASGFYFG DFGMS WVRQVPG 78 amino acid KGLEWVSGIDWSGRSTGYVDSMKG RLTISRDNDKSSLYLQ MNDLRGEDTAVYYCARV GRLCSGDSCDSMGAFDLWGQGT MVTVSS 12G10 VLgacatccagttgacccagtctccttccaccctgtctgcatctataggagacagcgtcaccatcactt 79kappa-type gc cgggccagttcagaatattgataactggttggcctggtatcaacacagaaaccagggaaagccc DNActagactcctgatctacaaggcgtctagtctaggaagtggggtctcatcaaagttcagaggcagtggatttgggacagagttcactctcaccatcaccagcctgcagcctgatgactttgcaacctattattg tcagaagtctaatggctattctcgtact tttggccaggggaccaaagtggatatcaaa 12G10 VLDIQLTQSPSTLSASIGDSVTITCRASQNIDNWLAWYQQKPGKA 77 kappa-type PRLLIYKASSLGSGVSSKFRGSGFGTEFTLTITSLQPDDFATYY amino acid C QKSNGYSRT FGQGTKVDIK

Antibody Production and Purification

Recombinant monoclonal human antibodies are expressed upon transfectionof antibody-coding expression vectors into HEK293T or Chinese HamsterOvary cells by the Polyethylenimine transfection method (PEI,Polyscience Warrington, USA). After transfection, cells are cultured inreduced serum medium (Opti-MEM® I supplemented with GlutaMAX™-I, Gibco)for 3-6 days. Subsequently, supernatants are collected andIgG-antibodies are purified using protein A columns (GE HealthCare,Sweden) on a fast protein liquid chromatography device (FPLC) (AKTA, GEHealthCare, Sweden).

Example 1: Detection of Peanut-Secific Antibodies by ELISA Assay

Enzyme linked immunosorbent assay (ELISA) was used to evaluate allergenreactivity in the sera of allergic patients. Allergens (Ara h 2, Ara h1, Ara h 3, Ara h 6, peanut extract) were coated on plates andanti-peanut antibody levels in sera from allergic patients were detectedusing HRP-conjugated anti-human antibody. Altogether, sera from 84allergic patients, were used in the assays. The following protocoldescribes the experimental procedures for the detection of anti-allergenantibodies by ELISA assay. Allergic patients have shown seroreactivityagainst several Ara h peanut allergens, implicating the suitability ofthese antibodies against allergic response induced by several peanutallergens.

ELISA for the Detection of Peanut-Specific Antibodies

96 well microplates (Costar®, Corning Incorporated, Corning, N.Y., USA)were coated with peanut allergens either extracted and purified frompeanuts or recombinantly expressed. Major peanut allergens included inthe example are Ara h 1, Ara h 2, Ara h 3 and Ara h 6 (IndoorBiotechnologies, Cardiff, UK). Plates were washed with PBS-Tween 0.05%and blocked lh at room temperature with PBS containing 5% Milk(Rapilait, Migros, Zurich, Switzerland) or 2% bovine serum albumin (BSA,Sigma-Aldrich Chemie GmbH, Buchs, Switzerland). Patient sera, B cellconditioned medium, or recombinant antibody preparations were incubatedfor 2h at room temperature. Binding of human IgG4, IgE or total IgG tothe antigen of interest was determined using a HRP-conjugated anti humanantibody (anti-IgG-HRP from Jackson ImmunoResearch, West Grove, Pa.,USA; anti-IgE-HRP from Abcam, Cambridge, UK anti-IgG4-HRP fromThermoFisher Scientific, Carlsbad, Calif., USA) followed by measurementof the HRP activity using a tetramethylbenzidine substrate solution(TMB, Sigma-Aldrich Chemie GmbH, Buchs, Switzerland). For the resultssee FIG. 1-2.

Example 2: EC50 ELISA Determination of the Antibodies of the PresentInvention

EC50 binding of exemplary anti-peanut antibodies of the presentinvention to peanut allergens or peanut extract, was determined byELISA. Serial dilutions of MAbs (from 1000 ng/ml down to 0.0169 ng/ml)were incubated for 2 hours with antigen-coated plates (coating overnightat 4° C. or 1 h at 37° C. with 1 μg/ml antigen in PBS, followed by washout and blocking with 2% BSA in PBS). The plates were subsequentlywashed and binding of MAbs was detected with anti-human HRP-conjugatedFc-gamma-specific secondary antibody (Jackson ImmunoResearch, WestGrove, Pa., USA). Concentrations of MAb resulting in half of maximalbinding to respective antigens (EC50, ng/ml) were calculated usingGraphPad Prism 5 software on sigmoidal dose-response curves (variableslope, 4 parameters) obtained by plotting the log of the concentrationversus OD450 nm measurements; for the results see FIG. 3 and table 2.

TABLE 2 EC50 ELISA determination of binding of exemplary humananti-peanut allergen antibodies and murinized versions of the antibodiesdepicted in FIG. 3. EC50 [ng/ml], peanut allergens MY006 peanut otherallergens antibodies nArah2 extract nArah1 nArah3 nArah6 rArah8 rArah9nBetv1 rAna3o nCora9 h12G10 6.8 8.1 242.2 33.7 17.2 n.b. n.b. n.b. n.b.n.b. hm12G10 6.7 8.5 194.7 36.8 27.8 n.p. n.p. n.p. n.p. n.p. h37D5 5.66.5 263.8 65.2 22.1 n.b. n.b. n.b. n.b. n.b. hm37D5 5.2 7.1 115.1 72.334.0 n.p. n.p. n.p. n.p. n.p. h32B10 9.2 4.8 8.8 4.9 7.0 n.b. n.b. n.b.n.b. n.b. hm32B10 9.9 n.p. n.p. n.p. n.p. n.p. n.p. n.p. n.p. n.p. h2F85.6. 2.8 8.5 2.8 3.4 n.b. n.b. n.b. n.b. n.b. hm2F8 8.7 n.p. n.p. n.p.n.p. n.p. n.p. n.p. n.p. n.p. h7G6 4.7 2.8 1.9 1.3 2.6 n.b. n.b. n.b.n.b. n.b. hm 7G6 n.p. n.p. n.p. n.p. n.p. n.p. n.p. n.p. n.p. n.p. h4B26.7 n.p. 5.9 n.p. 7.0 n.b. n.b. n.b. n.b. n.b. hm4B2 10.2 n.p. n.p. n.p.n.p. n.p. n.p. n.p. n.p. n.p. n.b. = not binding, n.p. = experiment notperformed.

Example 3: Cross-Competition with Human-Mouse Chimeric Constructs ofAnti-Peanut Antibodies

As first step of mapping, differential binding of anti-peanut MAbs ofthe present invention to distinct antigen binding sites was examined todetermine the number of different binding sites. For this purpose, MAbswere expressed either with human (hMAb) or mouse (hmMab) Fc andcross-competition experiments were carried out by coating antigen onplates and by detecting binding of human Mabs in the presence oftitrated amount of hmMabs. Detection of hMAbs bound to the ligand wasperformed by a HRP conjugated secondary antibody directed against the Fcportion of the primary antibody (Jackson ImmunoResearch, West Grove,Pa., USA). As may be seen from Table 3, exemplary anti-peanut allergensantibodies 37D5 and 12G10 of the present invention compete each otherfor binding of Ara h 2 but not with antibodies 4B2 and 32B10, indicatingthat 12G10 and 37D5 bind other sites of Ara h 2 than 4B2 and 32B10.

TABLE 3 The differential binding of exemplary human monoclonalantibodies (hMAbs) of the present invention with the respective MAbswith mouse Fc (hmMAbs) to distinct binding sites was investigated incross-competition experiments with titrated hmMAbs (0-12,000 ng/ml) andanti-human IgG horseradish peroxidase (HRP)-conjugated detectionantibody. “n.c.”: non- competing antibodies; “competing”: competingantibodies. 37D5 32B10 12G10 4B2 37D5 competing n.c. competing n.c.32B10 n.c. competing n.c. n.c. 12G10 competing n.c. competing n.c. 4B2n.c. n.c. n.c. competing

Example 4: ELISA Inhibition Assay of Allergic Patient IgE Binding toAllergen

Microplates were coated with 0.05 m/ml of Ara h 1, Ara h 2, Ara h 3, orAra h 6, overnight at 4° C. After washing with PBS-Tween 0.05% andblocking with 2% BSA, plates were incubated with increasingconcentrations of peanut human IgG antibodies for 1 h at roomtemperature. Subsequently, plates were incubated with different seradilutions or different concentrations of murine antibody (as positivecontrol). IgE levels were detected using anti-human HRP-conjugatedFc-epsilon-specific secondary antibody (Abcam, Cambridge, UK) followedby addition of TMB solution (TMB, Sigma, Buchs, Switzerland). Binding ofmurine antibodies was detected using goat anti-mouse IgG (H+L)-HRP(Jackson ImmunoResearch, West Grove, Pa., USA). Results are shown intable 4.

TABLE 4 ELISA competition assay shows inhibition of allergic patient IgEbinding to allergen (Ara h 2). 37D5 and 12G10 antibody blocked thebinding of allergic patients' polyclonal IgE to Ara h 2 up to 60%.Antibody cocktail containing 32B10, 37D5, 4B2 and 12G10 blocked up to80% of polyclonal IgE from allergic patients. 32B10 12G10 37D5 4B2cocktail Patient A 27.76 ± 4.85 13.82 ± 5.31 27.77 ± 3.58 39.66 ± 5.9566.21 ± 0.6  Patient B 26.66 ± 1.63 31.16 ± 5.92 29.61 ± 1.33 30.65 ±2.48 58.75 ± 1.49 Patient C 21.00 ± 5.17 53.81 ± 3.35 55.08.42 ± 1.98   13.34 ± 5.71 64.62 ± 3.41 Patient D  8.50 ± 7.06 35.74 ± 7.96 35.51 ±4.53  9.23 ± 3.17 71.02 ± 1.48 Patient E 29.32 ± 2.03 36.24 ± 3.25 29.30± 3.05 21.91 ± 5.23 73.18 ± 5.29 Patient F 23.15 ± 4.81 45.34 ± 3.1036.25 ± 3.27 30.77 ± 0.73 75.06 ± 6.91 Patient G 32.83 ± 6.33 25.74 ±7.00  45.32 ± 17.11 31.77 ± 2.00 80.15 ± 5.73 Healthy donor 0.00 0.00 2.27 ± 64.04 0.00 0.00

Example 5: Basophil Activation Test. Human-Derived Anti-PeanutMonoclonal Antibodies Neutralize Natural Ara h 2 and Peanut ExtractMediated Basophil Activation

Basophil activation test was performed using whole blood from allergicpatients, sampled in Heparin tubes (S-Monovette® 02.1065, Sarstedt,Niimbrecht, Germany). To test the neutralizing capacity of antibodies,the optimal concentration of Ara h 2 or peanut extract was preincubatedwith one or serial dilutions of serum, supernatant or purified antibodysamples (one or a combination of them). Afterwards, 100 μl of wholeblood were stimulated with either natural Ara h 2 (10 ng/ml, IndoorBiotechnologies, Cardiff, UK) or peanut extract (10 ng/ml, IndoorBiotechnologies, Cardiff, UK) or stimulation buffer (negative control)in the presence/absence of different anti-peanut allergen antibodies orthe combination thereof (MY006 cocktail) or isotype control for 30minutes at 37° C., 5% CO2. Cells were stained simultaneously withanti-CCR3-APC, anti-CD203c-PE and anti-CD63-FITC (Biolegend, San Diego,Calif., USA). Basophils were gated as SSClow, CCR3high lymphocytes. Atleast 500 basophils were acquired using FACSCalibur (Becton DickinsonAG, Allschwil, Switzerland). Activation of basophils was quantifiedusing % of CD63+ CCR3+ basophils and/or median fluorescence intensity ofCD203c CCR3+ basophils. Data was analyzed using FlowJo Software (FlowJo,Ashland, Oreg., USA). Results are shown in FIG. 4.

Example 6: Epitope Mapping of Exemplary Peanut-Specific Antibodies

A competition ELISA was used to evaluate binding of exemplaryanti-peanut antibodies of the present invention to linear Ara h 2epitopes. Serial dilutions of linear Ara h 2 epitopes (100 μM down to0.19 nM; peptides with a length of 20 amino acids and an overlap of 12amino acids between the peptides) were pre-incubated with exemplaryanti-peanut antibodies (66 μM) for 1 hour at room temperature. Thepre-incubated antibody-epitope mixtures were subsequently incubated onplates coated with full length Ara h 2 (coating overnight at 4° C. with0.05 μg/ml Ara h 2 in PBS, followed by wash out and blocking with 2% BSAin PBS). The plates were subsequently washed and binding of MAbs to Arah 2 was detected HRP-conjugated goat anti human Fc-gamma-specificantibody (Jackson ImmunoResearch, West Grove, Pa., USA) followed bymeasurement of the HRP activity using a tetramethylbenzidine substratesolution (TMB, Sigma-Aldrich Chemie GmbH, Buchs, Switzerland). As may beseen from Table 5, exemplary anti-peanut allergen antibody 32B10 bindsto different linear Ara h 2 epitopes than 12G10 and 37D5. 4B2,7G6 and2F8 do not bind to any of the 20 linear epitopes suggesting binding toconformational epitopes.

TABLE 5Epitope binding of exemplary human monoclonal antibodies of the present invention was evaluated in a competition ELISA with serial dilutions oflinear Ara h 2 peptides (100 μM down to 0.19 nM). Antibodies 12G10 and 37D5 are binding peptides 8-11, antibody 32B10 binds to peptide 4. Antibodies 4B2, 7G6 and 2F8 do not bind any of the linear Ara h 2 peptides.SEQ ID Sequence/ description of pool 32B10 37D5 12G10 4B2 7G6 2F8 POOLCombination of peptides 1 to 5 Binding n.b n.b n.b. n.b. n.b.  81Peptide 1 ¹AMALKLTILVALALFLLAAH²⁰ n.b n.b. n.b. n.b. n.b. n.b.  82Peptide 2 ⁹ALALFLLAAHASARQQWELQ²⁸ n.b. n.b. n.b. n.b. n.b. n.b.  83Peptide 3 ¹⁷AHASARQQWELQGDRRCQSQ³⁶ n.b. n.b n.b n.b. n.b. n.b.  84Peptide 4 ²⁵WELQGDRRCQSQLERANLRP⁴⁴ Binding n.b. n.b. n.b. n.b. n.b.  85Peptide 5 ³³CQSQLERANLRPCEQHLMQK⁵² n.b. n.b. n.b. n.b. n.b. n.b. POOL 2Combination of peptides 6 to 10 n.b. Binding Binding n.b. n.b. n.b.  86Peptide 6 ⁴¹NLRPCEQHLMQKIQRDEDSY⁶⁰ n.b. n.b n.b n.b. n.b. n.b.  87Peptide 7 ⁴⁹LMQKIQRDEDSYGRDPYSPS⁶⁸ n.b. n.b. n.b. n.b. n.b. n.b.  88Peptide 8 ⁵⁷EDSYGRDPYSPSQDPYSPSQ⁷⁶ n.b. Binding Binding n.b. n.b. n.b. 89 Peptide 9 ⁶⁵YSPSQDPYSPSQDPDRRDPY⁸⁴ n.b Binding Binding n.b. n.b.n.b.  90 Peptide 10 ⁷³SPSQDPDRRDPYSPSPYDRR⁹² n.b. Binding Binding n.b.n.b. n.b. POOL 3 Combination of peptides  n.b. Binding Binding n.b. n.b.n.b. 11 to 15  91 Peptide 11 ⁸¹RDPYSPSPYDRRGAGSSQHQ¹⁰⁰ n.b BindingBinding n.b. n.b. n.b.  92 Peptide 12 ⁸⁹YDRRGAGSSQHQERCCNELN¹⁰⁸ n.b.n.b. n.b. n.b. n.b. n.b.  93 Peptide 13 ⁹⁷SQHQERCCNELNEFENNQRC¹¹⁶ n.b.n.b. n.b. n.b. n.b. n.b.  94 Peptide 14 ¹⁰⁵NELNEFENNQRCMCEALQQI¹²⁴ n.b.n.b. n.b. n.b. n.b. n.b.  95 Peptide 15 ¹¹³NQRCMCEALQQIMENQSDRL¹³² n.b.n.b. n.b. n.b. n.b. n.b. POOL 4 Combination of peptides  n.b. n.b. n.b.n.b. n.b. n.b. 16 to 20  96 Peptide 16 ¹²¹LQQIMENQSDRLQGRQQEQQ¹⁴⁰ n.b.n.b. n.b. n.b. n.b. n.b.  97 Peptide 17 ¹²⁹SDRLQGRQQEQQFKRELRNL¹⁴⁸ n.b.n.b. n.b. n.b. n.b. n.b.  98 Peptide 18 ¹³⁷RQQEQQFKRELRNLPQQCGLRA¹⁵⁶n.b. n.b. n.b. n.b. n.b. n.b.  99 Peptide 19 ¹⁴⁵LRNLPQQCGLRAPQRCDLEV¹⁶⁴n.b. n.b. n.b. n.b. n.b. n.b. 100 Peptide 20 ¹⁵³GLRAPQRCDLEVESGGRDRY¹⁷²n.b. n.b. n.b. n.b. n.b. n.b. “n.b.”: not binding to peptide; “Binding”:binding to peptide.

Example 7: Validation of Peanut Antibodies in Animals

Mice C3H/HeJ, BALB/c, C57BL/6 and/or AKR/J are sensitized byintragastric (i.g.) or intraperitoneal (i.p.) administration of peanut(e.g. ground whole peanut, crude peanut extracts or whole peanutextracts) and adjuvant (e.g. cholera toxin, alum) on a weekly basis forup to 12 weeks. Control mice receive PBS with selected adjuvant. After2-3 months of sensitization, mice are challenged by i.g. or i.p.administration of peanut (e.g. ground whole peanut, crude peanutextracts or whole peanut extracts). Treatment consists of i.p. orintravenous (i.v.) administration of different antibody doses priorchallenge.

At several time points during the course of the model, blood is takenfor the measurement of antibodies (total and peanut specific IgE and IgGantibodies), mMCP-1 (mast cell protease 1), histamine and leukotrienelevels.

Anaphylactic symptoms are evaluated 30 minutes after the challenge doseutilizing a defined scoring system (see table below). Scoring ofsymptoms is performed in a blinded manner by two independentinvestigators. After euthanization, ear, spleen and blood samples arecollected. Blood samples are analyzed as described above. Histologicalsections are prepared and stained with toluidine blue or Giemsa stain.Degranulated mast cells are toluidine blue- or Giemsa positive with 5 ormore stained granules completely outside the cell. 400 mast cells areclassified.

Anaphylaxis scoring system (ref Liu et al. 2016 review or originalpapers):

Score Anaphylactic symptoms 0 no symptoms 1 scratching and rubbingaround the nose and head 2 puffiness around the eyes and mouth, pilarerecti (erection of the hair of the skin), reduced activity and/ ordecreased activity with increased respiratory rate 3 wheezing, laboredrespiration, and cyanosis around the mouth and tail 4 slight or noactivity after prodding, or tremor and convulsion 5 death

Antibody effects are evaluated by comparing clinical reactions inantibody treated and control animals using the anaphylaxis scoringsystem (illustrated in the table above). Therapeutic efficacy ofanti-peanut antibodies disclosed herein is indicated by a reduction orabsence of anaphylactic symptoms in antibody treated animals compared tocontrol animals.

Alternatively or in addition, antibody effects are evaluated bycomparing serum peanut specific IgE antibodies, mMCP-1 levels and/orplasma histamine and leukotriene levels in antibody treated and controlanimals using ELISA-based assays. Therapeutic efficacy of anti-peanutantibodies disclosed herein is indicated by a reduction or absence ofserum peanut-specific IgE antibodies and/or plasma histamine in antibodytreated animals relative to control animals. Alternatively or inaddition, antibody effects are evaluated by comparing cytokineexpression levels in challenged splenocytes derived from antibodytreated and control animals.

Example 8: Leukotriene Release Assay

Leukotriene release assay was performed using whole blood from allergicpatients, sampled in Heparin tubes (S-Monovette® 02.1065, Sarstedt,Niimbrecht, Germany). Leukocytes were isolated by dextran sedimentation.To test the neutralizing capacity of antibodies, titrations of peanutextract (0-30 ng/ml) were preincubated with purified antibody samples(one or a combination of them, 1.2 μg/ml) for 1 h at room temperature.Afterwards, leukocytes were stimulated with titrated peanut extract(Indoor Biotechnologies, Cardiff, UK) or stimulation buffer (negativecontrol) in the presence/absence of different anti-peanut allergenantibodies or the combination thereof (MY006 cocktail) or isotypecontrol for 40 minutes at 37° C., 5% CO2. Supernatant was collected andactivation of basophils was quantified using CAST® ELISA (Balmann,Schonenbuch, Switzerland). See FIG. 5 showing that human-derivedanti-peanut monoclonal antibodies neutralize peanut extract mediatedbasophil activation.

Example 9: Passive Sensitization Leukotriene Release Assay

Passive sensitization leukotriene release assay was performed usingwhole blood from healthy donors, sampled in Heparin tubes. Leukocyteswere isolated by dextran sedimentation. Surface IgE was removed bystripping with lactic acid, pH 3.9 (8 minutes 4° C.). Leukocytes werere-sensitized by incubation with plasma from allergic patients (1 h at37° C.). To test the neutralizing capacity of antibodies, titrations ofpeanut extract (0-30 ng/ml) were preincubated with purified antibodysamples (one or a combination of them, 1.2 μg/ml). Afterwards,re-sensitized leukocytes were stimulated with titrated peanut extract(Indoor Biotechnologies, Cardiff, UK) or stimulation buffer (negativecontrol) in the presence/absence of different anti-peanut allergenantibodies or the combination thereof (MY006 cocktail) or isotypecontrol for 40 minutes at 37° C., 5% CO2. Supernatant was collected andactivation of basophils was quantified using CAST® ELISA (Balmann,Schonenbuch, Switzerland). See FIG. 6 showing that human-derivedmonoclonal antibodies decrase peanut extract mediated basophilactivation.

The application further contains the following embodiments:

Embodiment 1. Antibody or binding fragment thereof capable of binding toa food allergen.

Embodiment 2. Antibody or binding fragment thereof according toembodiment 1, wherein the antibody is human-derived.

Embodiment 3. Antibody or binding fragment thereof according toembodiment 1 or 2, wherein the food allergen is a peanut allergen.

Embodiment 4. Antibody or binding fragment thereof according toembodiment 3, wherein the peanut allergen is selected from the groupconsisting of Ara h 1, Ara h 2, Ara h 3, Ara h 4, Ara h 5, Ara h 6/7,Ara h 8, Ara h 9 and Ara h 10/11.

Embodiment 5. Antibody or binding fragment thereof according toembodiment 4, wherein the peanut allergen is selected from the groupconsisting of Ara h 1, Ara h 2, Ara h 3 and Ara h 6, or a combinationthereof.

Embodiment 6. Antibody or binding fragment thereof according to any oneof embodiments 1 to 5, wherein the antibody is capable of binding atleast one of the Ara h2 epitopes selected from the group consisting ofthe amino acid sequences set forth in SEQ ID NOs: 84, 88, 89, 90 and 91.

Embodiment 7. Antibody or binding fragment thereof according to any oneof embodiments 3 to 5, wherein the peanut allergen is of peanut origin,recombinantly expressed or is a synthetic peanut peptide.

Embodiment 8. Antibody or binding fragment thereof according to any oneof the preceding embodiments, wherein the antibody is a monoclonalantibody and/or wherein the antibody is a recombinant antibody.

Embodiment 9. Antibody or binding fragment thereof according to any oneof the preceding embodiments, wherein the antibody is an IgG or IgAantibody.

Embodiment 10. Antibody or binding fragment thereof according to any oneof the preceding embodiments, wherein the variable regions, portionsthereof or the CDRs are human-derived.

Embodiment 11. Antibody or binding fragment thereof according to any oneof the preceding embodiments, wherein the variable regions, portionsthereof or the CDRs are derived from an IgE antibody and grafted in ascaffold of an IgG or IgA antibody.

Embodiment 12. Antibody or binding fragment thereof according to any oneof the preceding embodiments, wherein the peptide sequence of theantibody or binding fragment thereof is identical or at least 60%identical to the sequence of the antibody extracted from the human.

Embodiment 13. Antibody or binding fragment thereof according to any oneof embodiment 2 to 12, wherein the human is selected from the group of ahuman suffering from peanut allergy, a peanut-sensitized human withoutclinical relevant allergy, a human suffering from peanut allergy thatunderwent immunotherapy, a human that has outgrown peanut allergy and ahuman of unknown clinical history for peanut allergy.

Embodiment 14. Antibody or binding fragment thereof according to any oneof the preceding embodiments, wherein the antibody has an EC50 of atmost 270 ng/ml, preferably at most 70 ng/ml, at most 40 ng/ml, at most25 ng/ml, at most 15 ng/ml, at most 4.9 ng/ml, at most 1.3 ng/ml for atleast one of the peanuts allergens selected from the group consisting ofAra h 2, Ara h 1, Ara h 3 and Ara h 6.

Embodiment 15. Antibody or binding fragment thereof according to any oneof the preceding embodiments, wherein the antibody has an EC50 of atmost 10 ng/ml, preferably at most 7 ng/ml, more preferably at most 4.8ng/ml, most preferably 2.8 ng/ml for peanut extract.

Embodiment 16. Antibody or binding fragment thereof according to any oneof the preceding embodiments, wherein the antibody is capable ofreducing, inhibiting or neutralizing allergen-mediated biologicalactivity.

Embodiment 17. Antibody or binding fragment thereof according to any oneof the preceding embodiments, wherein the antibody is capable ofreducing or inhibiting the binding of an IgE antibody to the foodallergen.

Embodiment 18. Antibody or binding fragment thereof according to any ofthe preceding embodiments, comprising a light chain variable regionand/or a heavy chain variable region, wherein the light chain variableregion comprises a CDR1 set forth in SEQ ID No: 1 or sequences at least65% identical thereto, a CDR2 set forth in SEQ ID No: 2 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 3 orsequences at least 65% identical thereto; wherein the heavy chainvariable region comprises a CDR1 set forth in SEQ ID No: 4 or sequencesat least 65% identical thereto, a CDR2 set forth in SEQ ID No: 5 orsequences at least 65% identical thereto, and a CDR3 set forth in SEQ IDNo: 6 or sequences at least 65% identical thereto.

Embodiment 19. Antibody or binding fragment thereof according to any oneof embodiments 1 to 17, comprising a light chain variable region and/ora heavy chain variable region, wherein the light chain variable regioncomprises a CDR1 set forth in SEQ ID No: 15 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 16 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 17 orsequences at least 65% identical thereto; wherein the heavy chainvariable region comprises a CDR1 set forth in SEQ ID No: 18 or sequencesat least 65% identical thereto, a CDR2 set forth in SEQ ID No: 19 orsequences at least 65% identical thereto, a CDR3 set forth in SEQ ID No:20 or sequences at least 65% identical thereto.

Embodiment 20. Antibody or binding fragment thereof according to any oneof embodiments 1 to 17, comprising a light chain variable region and/ora heavy chain variable region, wherein the light chain variable regioncomprises a CDR1 set forth in SEQ ID No: 29 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 30 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 31 orsequences at least 65% identical thereto; wherein the heavy chainvariable region comprises a CDR1 set forth in SEQ ID No: 32 or sequencesat least 65% identical thereto, a CDR2 set forth in SEQ ID No: 33 orsequences at least 65% identical thereto, a CDR3 set forth in SEQ ID No:34 or sequences at least 65% identical thereto.

Embodiment 21. Antibody or binding fragment thereof according to any oneof embodiments 1 to 17, comprising a light chain variable region and/ora heavy chain variable region, wherein the light chain variable regioncomprises a CDR1 set forth in SEQ ID No: 43 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 44 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 45 orsequences at least 65% identical thereto; and/or wherein the heavy chainvariable region comprises a CDR1 set forth in SEQ ID No: 46 or sequencesat least 65% identical thereto, a CDR2 set forth in SEQ ID No: 47 orsequences at least 65% identical thereto, a CDR3 set forth in SEQ ID No:48 or sequences at least 65% identical thereto.

Embodiment 22. Antibody or binding fragment thereof according to any oneof embodiments 1 to 17, comprising a light chain variable region and/ora heavy chain variable region, wherein the light chain variable regioncomprises a CDR1 set forth in SEQ ID No: 57 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 58 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 59 orsequences at least 65% identical thereto; and/or wherein the heavy chainvariable region comprises a CDR1 set forth in SEQ ID No: 60 or sequencesat least 65% identical thereto, a CDR2 set forth in SEQ ID No: 61 orsequences at least 65% identical thereto, a CDR3 set forth in SEQ ID No:62 or sequences at least 65% identical thereto.

Embodiment 23. Antibody or binding fragment thereof according to any oneof embodiments 1 to 17, comprising a light chain variable region and/ora heavy chain variable region, wherein the light chain variable regioncomprises a CDR1 set forth in SEQ ID No: 71 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 72 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 73 orsequences at least 65% identical thereto; and/or wherein the heavy chainvariable region comprises a CDR1 set forth in SEQ ID No: 74 or sequencesat least 65% identical thereto, a CDR2 set forth in SEQ ID No: 75 orsequences at least 65% identical thereto, a CDR3 set forth in SEQ ID No:76 or sequences at least 65% identical thereto.

Embodiment 24. Antibody or binding fragment thereof according to any oneof embodiments 1 to 17, comprising a light chain variable region and/ora heavy chain variable region, wherein the light chain variable regioncomprises a CDR1 set forth in SEQ ID No: 1 or sequences at least 91%identical thereto, a CDR2 set forth in SEQ ID No: 2 or sequences atleast 71% identical thereto, a CDR3 set forth in SEQ ID No: 3; whereinthe heavy chain variable region comprises a CDR1 set forth in SEQ ID No:4 or sequences at least 80% identical thereto, a CDR2 set forth in SEQID No: 5 or sequences at least 82% identical thereto, a CDR3 set forthin SEQ ID No: 6 or a sequence at least 94% identical thereto.

Embodiment 25. Antibody or binding fragment thereof according to any oneof embodiments 1 to 17, comprising a light chain variable region and/ora heavy chain variable region, wherein the light chain variable regioncomprises a CDR1 set forth in SEQ ID No: 71 or sequences at least 91%identical thereto, a CDR2 set forth in SEQ ID No: 72 or sequences atleast 71% identical thereto, a CDR3 set forth in SEQ ID No: 73; whereinthe heavy chain variable region comprises a CDR1 set forth in SEQ ID No:74 or sequences at least 80% identical thereto, a CDR2 set forth in SEQID No: 75 or sequences at least 82% identical thereto, a CDR3 set forthin SEQ ID No: 76 or sequences at least 94% identical thereto.

Embodiment 26. Antibody or binding fragment thereof according to any oneof the preceding embodiments, comprising a CL and/or CH constant regioncomprising an amino acid sequence selected from the CL amino acidsequences SEQ ID NOS: 9, 23, 37, 51 and 65 or an amino acid sequencewith at least 60% identity and an amino acid sequence selected from theCH amino acid sequences SEQ ID NOS: 10, 24, 38, 52 and 66 or an aminoacid sequence with at least 60% identity.

Embodiment 27. Antibody or binding fragment thereof according to any oneof the preceding embodiments, wherein the antibody is selected from thegroup consisting of a full length antibody , a multispecific antibody, asingle chain Fv fragment (scFv), a F(ab′) fragment, a F(ab′)2 fragment,a F(ab)c fragment, a single domain antibody fragment (sdAB) and amultispecific antibody fragment.

Embodiment 28. Antibody or binding fragment thereof comprising a lightchain variable region and/or a heavy chain variable region, wherein thelight chain variable region comprises a CDR1 set forth in SEQ ID No: 1or sequences at least 65% identical thereto, a CDR2 set forth in SEQ IDNo: 2 or sequences at least 65% identical thereto, a CDR3 set forth inSEQ ID No: 3 or sequences at least 65% identical thereto; wherein theheavy chain variable region comprises a CDR1 set forth in SEQ ID No: 4or sequences at least 65% identical thereto, a CDR2 set forth in SEQ IDNo: 5 or sequences at least 65% identical thereto, and a CDR3 set forthin SEQ ID No: 6 or sequences at least 65% identical thereto.

Embodiment 29. Antibody comprising a light chain variable region and/ora heavy chain variable region, wherein the light chain variable regioncomprises a CDR1 set forth in SEQ ID No: 15 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 16 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 17 orsequences at least 65% identical thereto; wherein the heavy chainvariable region comprises a CDR1 set forth in SEQ ID No: 18 or sequencesat least 65% identical thereto, a CDR2 set forth in SEQ ID No: 19 orsequences at least 65% identical thereto, a CDR3 set forth in SEQ ID No:20 or sequences at least 65% identical thereto.

Embodiment 30. Antibody or binding fragment thereof comprising a lightchain variable region and/or a heavy chain variable region, wherein thelight chain variable region comprises a CDR1 set forth in SEQ ID No: 29or sequences at least 65% identical thereto, a CDR2 set forth in SEQ IDNo: 30 or sequences at least 65% identical thereto, a CDR3 set forth inSEQ ID No: 31 or sequences at least 65% identical thereto; wherein theheavy chain variable region comprises a CDR1 set forth in SEQ ID No: 32or sequences at least 65% identical thereto, a CDR2 set forth in SEQ IDNo: 33 or sequences at least 65% identical thereto, a CDR3 set forth inSEQ ID No: 34 or sequences at least 65% identical thereto.

Embodiment 31. Antibody or binding fragment thereof, comprising a lightchain variable region and/or a heavy chain variable region, wherein thelight chain variable region comprises a CDR1 set forth in SEQ ID No: 43or sequences at least 65% identical thereto, a CDR2 set forth in SEQ IDNo: 44 or sequences at least 65% identical thereto, a CDR3 set forth inSEQ ID No: 45 or sequences at least 65% identical thereto; and/orwherein the heavy chain variable region comprises a CDR1 set forth inSEQ ID No: 46 or sequences at least 65% identical thereto, a CDR2 setforth in SEQ ID No: 47 or sequences at least 65% identical thereto, aCDR3 set forth in SEQ ID No: 48 or sequences at least 65% identicalthereto.

Embodiment 32. Antibody or binding fragment thereof, comprising a lightchain variable region and/or a heavy chain variable region, wherein thelight chain variable region comprises a CDR1 set forth in SEQ ID No: 57or sequences at least 65% identical thereto, a CDR2 set forth in SEQ IDNo: 58 or sequences at least 65% identical thereto, a CDR3 set forth inSEQ ID No: 59 or sequences at least 65% identical thereto; and/orwherein the heavy chain variable region comprises a CDR1 set forth inSEQ ID No: 60 or sequences at least 65% identical thereto, a CDR2 setforth in SEQ ID No: 61 or sequences at least 65% identical thereto, aCDR3 set forth in SEQ ID No: 62 or sequences at least 65% identicalthereto.

Embodiment 33. Antibody or binding fragment thereof, comprising a lightchain variable region and/or a heavy chain variable region, wherein thelight chain variable region comprises a CDR1 set forth in SEQ ID No: 71or sequences at least 65% identical thereto, a CDR2 set forth in SEQ IDNo: 72 or sequences at least 65% identical thereto, a CDR3 set forth inSEQ ID No: 73 or sequences at least 65% identical thereto; and/orwherein the heavy chain variable region comprises a CDR1 set forth inSEQ ID No: 74 or sequences at least 65% identical thereto, a CDR2 setforth in SEQ ID No: 75 or sequences at least 65% identical thereto, aCDR3 set forth in SEQ ID No: 76 or sequences at least 65% identicalthereto.

Embodiment 34. Antibody or binding fragment thereof comprising a lightchain variable region and/or a heavy chain variable region, wherein thelight chain variable region comprises a CDR1 set forth in SEQ ID No: 1or sequences at least 91% identical thereto, a CDR2 set forth in SEQ IDNo: 2 or sequences at least 71% identical thereto, a CDR3 set forth inSEQ ID No: 3; wherein the heavy chain variable region comprises a CDR1set forth in SEQ ID No: 4 or sequences at least 80% identical thereto, aCDR2 set forth in SEQ ID No: 5 or sequences at least 82% identicalthereto, a CDR3 set forth in SEQ ID No: 6 or a sequence at least 94%identical thereto.

Embodiment 35. Antibody or binding fragment thereof comprising a lightchain variable region and/or a heavy chain variable region, wherein thelight chain variable region comprises a CDR1 set forth in SEQ ID No: 71or sequences at least 91% identical thereto, a CDR2 set forth in SEQ IDNo: 72 or sequences at least 71% identical thereto, a CDR3 set forth inSEQ ID No: 73; wherein the heavy chain variable region comprises a CDR1set forth in SEQ ID No: 74 or sequences at least 80% identical thereto,a CDR2 set forth in SEQ ID No: 75 or sequences at least 82% identicalthereto, a CDR3 set forth in SEQ ID No: 76 or sequences at least 94%identical thereto.

Embodiment 36. An antibody or binding fragment thereof which competeswith an antibody according to any one of embodiments 1 to 35 forspecific binding to the allergen.

Embodiment 37. A polynucleotide encoding the antibody or bindingfragment thereof according to any one of embodiments 1 to 36.

Embodiment 38. A vector comprising the polynucleotide of embodiment 37.

Embodiment 39. A cell comprising the polynucleotide of embodiment 37 orthe vector of embodiment 38.

Embodiment 40. A method for preparing an anti-allergen antibody orallergen-binding fragment thereof, consisting of culturing the cell ofembodiment 39 and isolating the antibody or allergen binding fragmentthereof from the cell or culture medium of the cell.

Embodiment 41. Antibody composition comprising at least two antibodies,wherein at least one of the antibodies is selected from the antibodiesas defined in embodiments 18 to 26 or 28 to 35.

Embodiment 42. Antibody composition according to embodiment 41, whereinat least two of the antibodies are selected from the antibodies asdefined in embodiments 18 to 26 or 28 to 35.

Embodiment 43. Antibody composition according to embodiment 42, whereinat least three of the antibodies are selected from the antibodies asdefined in embodiments 18 to 26 or 28 to 35.

Embodiment 44. A pharmaceutical composition comprising at least one ofthe compounds selected from the group consisting of antibody or bindingfragment of any one of embodiments 1 to 36, or the antibody compositionaccording to embodiments 41 to 43, the polynucleotide of embodiment 37,the vector of embodiment 38 and the cell of embodiment 39.

Embodiment 45. A pharmaceutical composition according to embodiment 44,comprising the antibody or binding fragment of any one of embodiments 1to 36 or the antibody composition according to embodiments 41 to 43.

Embodiment 46. A pharmaceutical composition according to embodiments 44to 45, further comprising a pharmaceutical acceptable carrier.

Embodiment 47. A pharmaceutical composition according to embodiments 44to 46, wherein the pharmaceutical composition further comprises at leastone additional agent useful for treating peanut allergy.

Embodiment 48. A pharmaceutical composition according to embodiment 47wherein the additional agent useful for treating peanut allergy isselected from the group 13-adrenergic agonists, epinephrine,antihistamine, corticosteroid, anti-IgE antibody, anti-IgE antibodybinding fragment, peptide vaccine and further antibodies capable ofbinding to a peanut allergen.

Embodiment 49. A pharmaceutical composition according to embodiments 44to 48, wherein the composition is intended for subcutaneous,intravenous, intramuscular, intraperitoneally intranasal and/orinhalative administration.

Embodiment 50. Kit comprising (i) the pharmaceutical compositionaccording to embodiments 44 to 46, (ii) at least one additional agentuseful for treating peanut allergy selected from the group 13-adrenergicagonist, antihistamine, corticosteroid, anti-IgE antibody, anti-IgEantibody binding fragment, peptide vaccine and further antibodiescapable of binding to a peanut allergen.

Embodiment 51. Kit according to embodiment 50, wherein the at least oneadditional agent is a β-adrenergic agonist, preferably epinephrine.

Embodiment 52. Antibody or binding fragment thereof according to any oneof embodiments 1 to 36, antibody composition according to embodiments 41to 43, polynucleotide according to embodiment 37, vector according toembodiment 38, cell according to embodiment 39 or pharmaceuticalcomposition according to embodiments 44 to 49 for use in the treatmentof peanut allergy.

Embodiment 53. Antibody or binding fragment, antibody composition,polynucleotide, vector, cell or pharmaceutical composition for use inthe treatment of peanut allergy according to embodiment 52, wherein thetreatment is prophylactic or therapeutic.

Embodiment 54. A method of evaluating the capacity of a candidateantibody or binding fragment thereof to inhibit allergen binding/and/orallergen-induced activity in a human, wherein the method comprises

(i) incubating the candidate antibody or binding fragment thereof with acomposition comprising IgEs derived from the human and a food allergen;

(ii) evaluating whether the candidate antibody inhibits allergen bindingto IgEs/and/or allergen-induced activity in the composition comprisingIgEs derived from the human.

Embodiment 55. The method of embodiment 54, further comprising the stepof

(iii) determining whether the administration of the candidate antibodyis a suitable treatment for a patient suffering from food allergy basedon the result of step (ii).

Embodiment 56. Method according to embodiments 54 and 55, wherein thecandidate antibody is an antibody as defined in embodiments 1 to 36.

Embodiment 57. Method according to embodiments 54 to 56, wherein in step(i) the composition comprising IgEs from the human comprises basophils.

Embodiment 58. Method according to embodiments 54 to 57, wherein thebasophils are derived from an allergic patient.

Embodiment 59. Method according to embodiments 54 to 58, wherein thebasophils are donor-derived IgE-stripped basophils.

Embodiment 60. Method according to embodiments 54 to 59, wherein theexpression of CD63+ at the surface of the basophil and/or the expressionof CD203c is measured.

Embodiment 61. Method according to embodiments 54 to 60, wherein thebasophils are identified based on the CCR3 expression.

Embodiment 62. Method according to embodiments 54 to 59, wherein thesecretion of a mediator from basophils is measured.

Embodiment 63. Method according to embodiment 62, wherein the mediatoris leukotriene.

Embodiment 64. Method according to embodiment 63, wherein the mediatoris sulfidoleukotrine.

Embodiment 65. Method according to any one of embodiments 54 to 64,wherein the composition comprising IgEs derived from the human isplasma, sera, blood, saliva, peripheral blood mononuclear cell (PBMC),leukocytes, basophils or IgEs stripped from basophils.

Embodiment 66. Method according to any one of embodiments 54 to 65,wherein the human is suffering from food allergy.

Embodiment 67. Method according to any one of embodiments 54 to 66,wherein the human is suffering from peanut allergy.

Embodiment 68. Method according to any one of embodiments 54 to 67,wherein the food allergen is a peanut allergen.

Embodiment 69. Method according to embodiment 68, wherein the peanutallergen is selected from the group consisting of Ara h 1, Ara h 2, Arah 3, Ara h 4, Ara h 5, Ara h 6/7, Ara h 8, Ara h 9 and Ara h 10/11.

Embodiment 70. Method according to embodiment 69, wherein the peanutallergen is selected from the group consisting of Ara h 1, Ara h 2, Arah 3 and Ara h 6, or a combination thereof.

Embodiment 71. Method according to any one of embodiment 68 to 70,wherein the peanut allergen is recombinantly expressed, syntheticallygenerated or is of peanut origin.

Embodiment 72. A method of detecting or quantifying whether an allergenis present in a sample comprising the following steps:

i) incubation of the sample with an antibody according to any one ofembodiments 1 to 36 or with an antibody composition according toembodiments 41 to 43,

ii) detecting the antibody which is bound to allergen in the sample.

Embodiment 73. The method of embodiment 72, wherein the antibody isdetectably labeled.

Embodiment 74. The method of embodiment 72, wherein the antibody isunlabeled and used in combination with a second antibody that isdetectably labeled.

Embodiment 75. The method of embodiment 73 or 74, wherein the detectablelabel is selected from the group consisting of an enzyme, aradioisotope, a fluorophore, a peptide and a heavy metal.

1. Antibody or binding fragment thereof capable of binding to a foodallergen.
 2. Antibody or binding fragment thereof according to claim 1,wherein the antibody is human-derived.
 3. Antibody or binding fragmentthereof according to claim 1 or 2, wherein the food allergen is a peanutallergen.
 4. Antibody or binding fragment thereof according to claim 3,wherein the peanut allergen is selected from the group consisting of Arah 1, Ara h 2, Ara h 3, Ara h 4, Ara h 5, Ara h 6/7, Ara h 8, Ara h 9 andAra h 10/11.
 5. Antibody or binding fragment thereof according to anyone of embodiments 1 to 4, wherein the antibody is capable of binding atleast one of the Ara h2 epitopes selected from the group consisting ofthe amino acid sequences set forth in SEQ ID NOs: 84, 88, 89, 90 and 91.6. Antibody or binding fragment thereof according to any one of thepreceding claims, wherein the antibody is a monoclonal antibody and/or arecombinant antibody.
 7. Antibody or binding fragment thereof accordingto any one of the preceding claims, wherein the antibody is an IgG orIgA antibody.
 8. Antibody or binding fragment thereof according to anyone of the preceding claims, wherein variable regions, portions thereofor the CDRs are derived from an IgE antibody and grafted in a scaffoldof an IgG or IgA antibody.
 9. Antibody or binding fragment thereofaccording to any one of claims 2 to 8, wherein the human is selectedfrom the group of a human suffering from peanut allergy, apeanut-sensitized human without clinical relevant allergy, a humansuffering from peanut allergy that underwent immunotherapy, a human thathas outgrown peanut allergy and a human of unknown clinical history forpeanut allergy.
 10. Antibody or binding fragment thereof according toany one of the preceding claims, wherein the antibody is capable ofreducing, inhibiting or neutralizing allergen-mediated biologicalactivity.
 11. Antibody or binding fragment thereof according to any oneof the preceding claims, wherein the antibody is capable of reducing orinhibiting the binding of an IgE antibody to the food allergen. 12.Antibody or binding fragment thereof according to any of the precedingclaims, wherein the antibody is selected from the group consisting ofthe following antibodies: a) antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 1 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 2 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 3 or sequences at least 65%identical thereto; wherein the heavy chain variable region comprises aCDR1 set forth in SEQ ID No: 4 or sequences at least 65% identicalthereto, a CDR2 set forth in SEQ ID No: 5 or sequences at least 65%identical thereto, and a CDR3 set forth in SEQ ID No: 6 or sequences atleast 65% identical thereto; b) antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 15 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 16 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 17 or sequences at least 65%identical thereto; wherein the heavy chain variable region comprises aCDR1 set forth in SEQ ID No: 18 or sequences at least 65% identicalthereto, a CDR2 set forth in SEQ ID No: 19 or sequences at least 65%identical thereto, a CDR3 set forth in SEQ ID No: 20 or sequences atleast 65% identical thereto; c) antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 29 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 30 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 31 or sequences at least 65%identical thereto; wherein the heavy chain variable region comprises aCDR1 set forth in SEQ ID No: 32 or sequences at least 65% identicalthereto, a CDR2 set forth in SEQ ID No: 33 or sequences at least 65%identical thereto, a CDR3 set forth in SEQ ID No: 34 or sequences atleast 65% identical thereto; d) antibody or binding fragment thereofcomprising a light chain variable region and/or a heavy chain variableregion, wherein the light chain variable region comprises a CDR1 setforth in SEQ ID No: 43 or sequences at least 65% identical thereto, aCDR2 set forth in SEQ ID No: 44 or sequences at least 65% identicalthereto, a CDR3 set forth in SEQ ID No: 45 or sequences at least 65%identical thereto; and/or wherein the heavy chain variable regioncomprises a CDR1 set forth in SEQ ID No: 46 or sequences at least 65%identical thereto, a CDR2 set forth in SEQ ID No: 47 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 48 orsequences at least 65% identical thereto; e) antibody or bindingfragment thereof comprising a light chain variable region and/or a heavychain variable region, wherein the light chain variable region comprisesa CDR1 set forth in SEQ ID No: 57 or sequences at least 65% identicalthereto, a CDR2 set forth in SEQ ID No: 58 or sequences at least 65%identical thereto, a CDR3 set forth in SEQ ID No: 59 or sequences atleast 65% identical thereto; and/or wherein the heavy chain variableregion comprises a CDR1 set forth in SEQ ID No: 60 or sequences at least65% identical thereto, a CDR2 set forth in SEQ ID No: 61 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 62 orsequences at least 65% identical thereto; and f) antibody or bindingfragment thereof comprising a light chain variable region and/or a heavychain variable region, wherein the light chain variable region comprisesa CDR1 set forth in SEQ ID No: 71 or sequences at least 65% identicalthereto, a CDR2 set forth in SEQ ID No: 72 or sequences at least 65%identical thereto, a CDR3 set forth in SEQ ID No: 73 or sequences atleast 65% identical thereto; and/or wherein the heavy chain variableregion comprises a CDR1 set forth in SEQ ID No: 74 or sequences at least65% identical thereto, a CDR2 set forth in SEQ ID No: 75 or sequences atleast 65% identical thereto, a CDR3 set forth in SEQ ID No: 76 orsequences at least 65% identical thereto.
 13. Antibody or bindingfragment thereof according to claim 12, wherein the antibody is selectedfrom the group consisting of the following antibodies: a) antibody orbinding fragment thereof comprising a light chain variable region and/ora heavy chain variable region, wherein the light chain variable regioncomprises a CDR1 set forth in SEQ ID No: 1, a CDR2 set forth in SEQ IDNo: 2, and a CDR3 set forth in SEQ ID No: 3, wherein the heavy chainvariable region comprises a CDR1 set forth in SEQ ID No: 4, a CDR2 setforth in SEQ ID No: 5, and a CDR3 set forth in SEQ ID No: 6; b) antibodyor binding fragment thereof comprising a light chain variable regionand/or a heavy chain variable region, wherein the light chain variableregion comprises a CDR1 set forth in SEQ ID No: 15, a CDR2 set forth inSEQ ID No: 16, and a CDR3 set forth in SEQ ID No: 17 or sequences atleast 65% identical thereto; wherein the heavy chain variable regioncomprises a CDR1 set forth in SEQ ID No: 18, a CDR2 set forth in SEQ IDNo: 19, and a CDR3 set forth in SEQ ID No: 20; c) antibody or bindingfragment thereof comprising a light chain variable region and/or a heavychain variable region, wherein the light chain variable region comprisesa CDR1 set forth in SEQ ID No: 29, a CDR2 set forth in SEQ ID No: 30,and a CDR3 set forth in SEQ ID No: 31; wherein the heavy chain variableregion comprises a CDR1 set forth in SEQ ID No: 32, a CDR2 set forth inSEQ ID No: 33, a CDR3 set forth in SEQ ID No: 34; d) antibody or bindingfragment thereof comprising a light chain variable region and/or a heavychain variable region, wherein the light chain variable region comprisesa CDR1 set forth in SEQ ID No: 43, a CDR2 set forth in SEQ ID No: 44,and a CDR3 set forth in SEQ ID No: 45; and/or wherein the heavy chainvariable region comprises a CDR1 set forth in SEQ ID No: 46, a CDR2 setforth in SEQ ID No: 47, and a CDR3 set forth in SEQ ID No: 48; e)antibody or binding fragment thereof comprising a light chain variableregion and/or a heavy chain variable region, wherein the light chainvariable region comprises a CDR1 set forth in SEQ ID No: 57, a CDR2 setforth in SEQ ID No: 58, and a CDR3 set forth in SEQ ID No: 59; and/orwherein the heavy chain variable region comprises a CDR1 set forth inSEQ ID No: 60, a CDR2 set forth in SEQ ID No: 61, and a CDR3 set forthin SEQ ID No: 62; and f) antibody or binding fragment thereof comprisinga light chain variable region and/or a heavy chain variable region,wherein the light chain variable region comprises a CDR1 set forth inSEQ ID No: 71, a CDR2 set forth in SEQ ID No: 72, and a CDR3 set forthin SEQ ID No: 73 or sequences at least 65% identical thereto; and/orwherein the heavy chain variable region comprises a CDR1 set forth inSEQ ID No: 74, a CDR2 set forth in SEQ ID No: 75, and a CDR3 set forthin SEQ ID No:
 76. 14. Antibody composition comprising at least twoantibodies, wherein at least one of the antibodies is selected from theantibodies as defined in claim
 12. 15. Pharmaceutical compositioncomprising at least one of the compounds selected from the groupconsisting of antibody or binding fragment of any one of claims 1 to 13,or the antibody composition according to claim
 14. 16. Pharmaceuticalcomposition according to claim 15, further comprising a pharmaceuticallyacceptable carrier.
 17. Kit comprising (i) the pharmaceuticalcomposition according to claim 15, (ii) at least one additional agentuseful for treating peanut allergy selected from the group β-adrenergicagonist, antihistamine, corticosteroid, anti-IgE antibody, anti-IgEantibody binding fragment, peptide vaccine and further antibodiescapable of binding to a peanut allergen.
 18. Kit according to claim 17,wherein the at least one additional agent is epinephrine.
 19. Antibodyor binding fragment thereof according to any one of claims 1 to 13,antibody composition according to claims 14 or pharmaceuticalcomposition according to claims 15 and 16, kit according to claims 17and 18 for use in the treatment of peanut allergy.
 20. Method ofevaluating the capability of a candidate antibody or binding fragmentthereof to inhibit allergen binding and/or allergen-induced activity ina human, comprising the following steps: (i) incubating the candidateantibody or binding fragment thereof together with basophils derivedfrom the human and a food allergen; (ii) measuring the level ofactivated basophils; wherein decreased levels of activated basophilscompared to a control indicate that the antibody is capable ofinhibiting allergen binding and/or allergen induced activity in a human.21. The method of claim 20, further comprising the step of (iii)determining whether the administration of the candidate antibody is asuitable treatment for a patient suffering from food allergy based onthe result of step (ii).
 22. Method according to claims 20 and 21,wherein the candidate antibody is an antibody as defined in claims 1 to14.
 23. Method according to claims 20 to 22, wherein in step (i) thecomposition comprising IgEs from the human comprises basophils. 24.Method according to claims 20 to 23, wherein the basophils are derivedfrom an allergic patient.
 25. Method according to claims 20 to 24,wherein the basophils are donor-derived IgE-stripped basophils. 26.Method according to claims 20 to 25, wherein the basophils aredonor-derived IgE-stripped basophils incubated with plasma from anallergy patient.
 27. Method according to claims 20 to 26, wherein thesecretion of a mediator from basophils is measured.
 28. Method accordingto claim 27, wherein the mediator is leukotriene.
 29. Method accordingto claim 28, wherein the mediator is sulfidoleukotrine.
 30. Methodaccording to any one of claims 20 to 29, wherein the compositioncomprising IgEs derived from the human is plasma, sera, blood, saliva,peripheral blood mononuclear cell (PBMC), leukocytes, basophils or IgEsstripped from basophils.
 31. Method according to claims 30, wherein thecomposition comprising IgEs derived from the human are leukocytes. 32.Method according to any one of claims 20 to 31, wherein the human issuffering from food allergy.
 33. Method according to any one of claims20 to 32, wherein the human is suffering from peanut allergy.
 34. Methodaccording to any one of claims 20 to 33, wherein the food allergen is apeanut allergen.
 35. Method according to claim 34, wherein the peanutallergen is selected from the group consisting of Ara h 1, Ara h 2, Arah 3, Ara h 4, Ara h 5, Ara h 6/7, Ara h 8, Ara h 9 and Ara h 10/11. 36.Method according to claim 35, wherein the peanut allergen is selectedfrom the group consisting of Ara h 1, Ara h 2, Ara h 3 and Ara h 6, or acombination thereof
 37. Method according to any one of claims 20 to 36,wherein the peanut allergen is recombinantly expressed, syntheticallygenerated or is of peanut origin.
 38. Method of claims 20 to 37, whereindecreased levels of activated basophils compared to the controlindicated that the antibody is suitable for the treatment of a patientsuffering from food allergy.
 39. Method according to any one of claims20 and 21, wherein the control comprises basophils derived from thehuman and a food allergen but does not contain the candidate antibody.40. A method of evaluating the capability of a candidate antibody orbinding fragment thereof to inhibit allergen binding, comprising thefollowing steps: (i) incubating the candidate antibody or bindingfragment thereof together with a composition comprising IgEs derivedfrom the human and a food allergen; (ii) measuring the level of IgEbound to antigen; wherein decreased levels of IgE bound to antigencompared to a control indicate that the antibody is capable ofinhibiting allergen binding.
 41. Method according to claim 40, whereindecreased levels of IgE bound to antigen compared to the controlindicated that the antibody is suitable for the treatment of a patientsuffering from food allergy.
 42. Method according to any one of claims40 and 41, wherein the control comprises IgEs derived from the human anda food allergen but does not contain the candidate antibody.
 43. Methodaccording to any one of claims 40 to 42, wherein the candidate antibodyis an antibody as defined in claims 1 to
 13. 44. A method of detectingor quantifying whether an allergen is present in a sample comprising thefollowing steps: i) incubation of the sample with an antibody accordingto any one of claims 1 to 13 or with an antibody composition accordingto claim 14, ii) detecting the antibody which is bound to allergen inthe sample.